Substituted heteroaryl compounds and methods of use

ABSTRACT

The present invention provides novel heteroaryl compounds, pharmaceutical acceptable salts and formulations thereof useful in preventing, treating or lessening the severity of a protein-mediated disease. The invention also provides pharmaceutically acceptable compositions comprising such compounds and methods of using the compositions in the treatment of protein kinase-mediated disease.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application Ser.No. 61/971,552, filed on Mar. 28, 2014, which is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

The present invention provides novel substituted aminopyrimidinecompounds, and salts thereof, which are useful in the treatment ofproliferative disease, autoimmune disease, allergic disease,inflammatory disease, transplantation rejection, and other diseases, inmammals. In particular, this invention relates to compounds thatmodulate the activity of JAK kinases, FLT3 kinase, and Aurora kinaseresulting in the modulation of inter- and/or intra-cellular signaling.This invention also relates to a method of using such compounds in thetreatment of proliferative disease, autoimmune disease, allergicdisease, inflammatory disease, transplantation rejection, and otherdiseases in mammals, especially humans, and to pharmaceuticalcompositions containing such compounds.

BACKGROUND OF THE INVENTION

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell. Protein kinases, containing asimilar 250-300 amino acid catalytic domain, catalyze thephosphorylation of target protein substrates. It is reported that manydiseases are associated with abnormal cellular responses triggered byprotein kinase-mediated events. These diseases include benign andmalignant proliferation disorders, diseases resulting from inappropriateactivation of the immune system, allograft rejection, graft vs hostdisease, autoimmune diseases, inflammatory diseases, bone diseases,metabolic diseases, neurological and neurodegenerative diseases, cancer,cardiovascular diseases, allergies and asthma, Alzheimer's disease andhormone-related diseases. Accordingly, there has been a substantialeffort in medicinal chemistry to find protein kinase inhibitors that areeffective as therapeutic agents.

The kinases may be categorized into families by the substrates in thephosphorylate (e.g., protein-tyrosine, protein-serine/threonine, lipids,etc.). Tyrosine phosphorylation is a central event in the regulation ofa variety of biological processes such as cell proliferation, migration,differentiation and survival. Several families of receptor andnon-receptor tyrosine kinases control these events by catalyzing thetransfer of phosphate from ATP to a tyrosine residue of specific cellprotein targets. Sequence motifs have been identified that generallycorrespond to each of these kinase families (Hanks et al., FASEB J.,1995, 9, 576-596; Knighton et al., Science, 1991, 253, 407-414;Garcia-Bustos et al., EMBO J., 1994, 13:2352-2361). Some non-limitingexamples of the protein kinase include abl, Aurora, Akt, bcr-abl, BIk,Brk, Btk, c-kit, c-Met, c-src, c-fms, CDK1, CDK2, CDK3, CDK4, CDK5,CDK6, CDK7, CDK8, CDK9, CDK10, cRafl, CSF1 R, CSK, EGFR, ErbB2, ErbB3,ErbB4, Erk, Fak, fes, Flt-3, FGFR1, FGFR2, FGFR3, FGFR4, FGFR5, Fgr,Flt-1, Fps, Frk, Fyn, Hck, IGF-1 R, INS-R, JAK, KDR, Lck, Lyn, MEK, p38,PDGFR, PIK, PKC, PYK2, ros, Tie, Tie-2, TRK, Yes, and Zap70.

Aurora kinase family is a collection of highly related serine/threoninekinase that are key regulators of mitosis, essential for accurate andequal segtion of genomic material from parent to daught cells. Membersof the Aurora kinase family include three related kinases kown asAurora-A, Aurora-B, and Aurora-C (also known as Aurora-1, Aurora-2, andAurora-3). Despite significant sequence homology, the localization andfunctions of these kinases are largely distinct from one another(Richard D. Carvajal, et al. Clin Cancer Res., 2006, 12(23): 6869-6875;Daruka Mahadevan, et al., Expert Opin. Drug Discov., 2007 2(7):1011-1026).

Aurora-A is ubiquitously expressed and regulates cell cycle eventsoccurring from late S phase through M phase, including centrosomematuration (Berdnik D, et al., Curr Biol., 2002, 12: 640-647), mitoticentry (Hirota T, et al., Cell, 2003, 114:585-598; Dutertre S, et al., JCell Sci., 2004, 117:2523-2531), centrosome separation (Marumoto T, etal. J Biol. Chem., 2003, 278:51786-51795), bipolar-spindle assembly(Kufer T A, et al. J Cell Biol., 2002; 158:617-623; Eyers P A, et al.,Curr Biol., 2003; 13:691-697), chromosome alignment on the metaphaseplate (Marumoto T, et al. J Biol Chem., 2003, 278:51786-51795; KunitokuN, et al., Dev Cell., 2003, 5: 853-864), cytokinesis (Marumoto T, etal., J Biol. Chem., 2003, 278:51786-51795), and mitotic exit. Aurora-Aprotein levels and kinase activity both increase from late G2 through Mphase, with peak activity in prometaphase. Once activated, Aurora-Amediates its multiple functions by interacting with various substratesincluding centrosomin, transforming acidic coiled-coil protein, cdc25b,Eg5, and centromere protein A.

Aurora-B is a chromosomal passenger protein critical for accuratechromosomal segregation, cytokinesis (Hauf S., et al. J Cell Biol.,2003, 161:281-94; Ditchfield C, et al., J Cell Biol., 2003, 161:267-80;Giet R, et al., J Cell Biol., 2001, 152:669-682; Goto H, et al., J Biol.Chem., 2003, 278:8526-8530), protein localization to the centromere andkinetochore, correct microtubule-kinetochore attachments (Murata-Hori M,et al., Curr. Biol., 2002, 12:894-899), and regulation of the mitoticcheckpoint. Aurora-B localizes first to the chromosomes during prophaseand then to the inner centromere region between sister chromatids duringprometaphase and metaphase (Zeitlin S G, et al. J Cell. Biol., 2001,155:11147-1157). Aurora-B participates in the establishment ofchromosomal biorientation, a condition where sister kinetochores arelinked to opposite poles of the bipolar spindle via amphitelicattachments. Errors in this process, manifesting as a merotelicattachment state (one kinetochore attached to microtubules from bothpoles) or a syntelic attachment state (both sister kinetochores attachedto microtubules from the same pole), lead to chromosomal instability andaneuploidy if not corrected before the onset of anaphase. The primaryrole of Aurora-B at this point of mitosis is to repair incorrectmicrotubule-kinetochore attachments (Hauf S, et al., J Cell Biol., 2003,161:281-294; Ditchfield C, et al., J Cell Biol., 2003, 161:267-280; LanW, et al. Curr Biol., 2004, 14:273-286). Without Aurora-B activity, themitotic checkpoint is compromised, resulting in increased numbers ofaneuploid cells, genetic instability, and tumorigenesis (Weaver B A, etal., Cancer Cell., 2005, 8:7-12).

Aurora-A overexpression is a necessary feature of Aurora-A-inducedtumorigenesis. In cells with Aurora-A overexpression, mitosis ischaracterized by the presence of multiple centrosomes and multipolarspindles (Meraldi P et al., EMBO J., 2002, 21:483-492.). Despite theresulting aberrant microtubule-kinetochore attachments, cells abrogatethe mitotic checkpoint and progress from metaphase to anaphase,resulting in numerous chromosomal separation defects. These cells failto undergo cytokinesis, and, with additional cell cycles, polyploidy andprogressive chromosomal instability develop (Anand S, et al., CancerCell, 2003, 3:51-62).

The evidence linking Aurora overexpression and malignancy prolifrationdisorders, such as colon, breast, lung, pancrease, prostate, bladder,head, neck, cervix, and ovarian cancers, liver, gastric and pancreatictumors, has stimulated interest in developing Aurora inhibitors forcancer therapy. In normal cells, Aurora-A inhibition results in delayed,but not blocked, mitotic entry, centrosome separation defects resultingin unipolar mitotic spindles, and failure of cytokinesis (Mammoto T, etal., J Biol. Chem., 2003, 278:51786-51795). Encouraging antitumoreffects with Aurora-A inhibition were shown in three human pancreaticcancer cell lines (Pane-1, MIA PaCa-2, and SU.86.86), with growthsuppression in cell culture and near-total abrogation of tumorigenicityin mouse xenografts (Hata T, et al., Cancer Res., 2005, 65:2899-2905).

Aurora-B inhibition results in abnormal kinetochore-microtubuleattachments, failure to achieve chromosomal biorientation, and failureof cytokinesis (Goto H, et al. J Biol Chem., 2003, 278:8526-30; SeversonAF₁ et al. Curr Biol., 2000, 10:1162-1171). Recurrent cycles of aberrantmitosis without cytokinesis result in massive polyploidy and,ultimately, to apoptosis (Hauf S, et al., J Cell Biol., 2003,161:281-294; Ditchfield C, et al., J Cell Biol, 2003, 161:267-80; GietR, et al., J Cell Biol., 2001, 152:669-682; Murata-Hori M, Curr. Biol.,2002, 12:894-899; Kallio et al., Curr Biol., 2002, 12:900-905).

Inhibition of Aurora-A or Aurora-B activity in tumor cells results inimpaired chromosome alignment, abrogation of the mitotic checkpoint,polyploidy, and subsequent cell death. These in vitro effects aregreater in transformed cells than in either non-transformed ornon-dividing cells (Ditchfield C, et al., J Cell Biol., 2003,161:267-280). Thus, targeting Aurora may achieve in vivo selectivity forcancer. Although toxicity to rapidly dividing cell of the hematopoieticand gastrointestinal system is expected, the activity and clinicaltolerability shown in xenograft models indicates the presence of areasonable therapeutic index. Given the preclinical antitumor activityand potential for tumor selectivity, several Aurora kinase inhibitorshave been developed.

FLT3 (Flt3, FMS-related tyrosine kinase 3), also known as FLK-2 (fetalliver kinase 2) and STK-1 (human stem cell kinase 1), belongs to amember of the class III receptor tyrosine kinase (RTK-III) family thatinclude KIT, PDGFR, FMS and FLT1 (Stirewalt D L, et al., Nat. Rev.Cancer, 2003, 3:650-665; Rosnet O, et al., Genomics 1991, 9:380-385;Yarden Y, et al., Nature, 1986, 323: 226-232; Stanley E R, et. al, J.Cell. Biochem., 1983, 21:151-159; Yarden Y, et al., EMBO J., 1987,6:3341-3351). FLT3 is a membrane-spanning protein and composed of fourdomains; an extracellular ligand-binding domains consisting of fiveimmunoglobin-like structures, a transmembrane (TM) domain, ajuxtamembrane (JM) domain and a cytoplasmic C-Terminal tyrosine kinase(TK) domain. (Agnes F. et al., Gene, 1994, 145:283-288, Scheijen B, etal., Oncogene, 2002, 21: 3314-3333).

The ligand for FLT3 (FLT3 or FL) was cloned in 1993 and shown to be aType I transmembrane protein expressed in cells of the hematopoieticbone marrow microenvironment, including bone marrow fibroblasts andother cells (Lyman S D, et al., Cell 1993, 75:1157-1167). Both themembrane-bound and soluable forms can activate the tyrosine kinaseactivity of the receptor and stimulate growth of progenitor cells in themarrow and blood. Binding of ligand to receptor induces dimerisation ofthe receptor and activation of the kinase domains; which thenautophosphorylate and catalyse phosphorylation of substrate proteins ofvarious signal transduction pathways such as signal transducer andactivator of transcription 5 (STAT5), RAS/mitogen-activated proteinkinase (RAS/MAPK), phosphoinositide 3-kinase (PI3K), src homologous andcollagen gene (SHC), SH2-containing inositol-5-phosphatase (SHIP), andcytoplasmic tyrosine phosphatase with 2 Src-homology 2 (SH2) domains(SHP2), which play important roles in cellular proliferation,differentiation, and survival (Dosil M, et al. Mol Cell Biol., 1993,13:6572-6585. Zhang S, Biochem Biophys Res Commun., 1999, 254:440-445).In addition to hemotopoietic cells, FLT3 gene is also expressed inplacenta, gonads and brain (Maroc N, et al. Oncogene, 1993, 8: 909-918)and also plays an importand role in the immune response (deLapeyriere O,et al., Leukemia, 1995, 9:1212-1218).

FLT3 has also been implicated in hematopoietic disorders which arepre-malignant disorders including myeloproliferative disorders, such asthrombocythemia, essential thrombocytosis (ET), myelofibrosis (MF),chronic idiopathic myelofibrosis (IMF), and polycythemia vera (PV),pre-malignant myelodysplastic syndromes. Hematological malignanciesinclude leukemias, lymphomas (non-Hodgkin's lymphoma), Hodgkin's disease(also called Hodgkin's lymphoma), and myeloma, for instance, acutelymphocytic leukemia (ALL), acute myeloid leukemia (AML), acutepromyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL),chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL).FLT3 is overexpressed at the levels in 70-100% of cases of acute myeloidleukemias (AML), and in a high percentage of T-acute lymphocyticleukemia (ALL) cases (Griffin J D, et al., Haematol J. 2004, 5:188-190), it is also overexpressed in a smaller subset of chronicmyeloid leukemia (CML) in blast crisis. Studies have shown that theleukemic cells of B lineage ALL and AML frequently co-express FL,setting up autocrine or paracrine signaling loops that result in theconstitutive activation of FLT3 (Zheng R, et. al., Blood., 2004, 103:267-274). A high level of the FLT3 ligand is found in the serum ofpatients with Langerhans cell histocytosis and systemic lupuserythematosus, which further implicates FLT3 signaling in thedysregulation of dendritic cell progenitors in those autoimmune diseases(Rolland et al., J. Immunol., 2005, 174:3067-3071).

Evidence is rapidly accumulating that many types of leukemias andmyeloproliferative syndromes have mutation in tyrosine kinases. FLT3mutations are one of the most frequent somatic alterations in AML,occurring in approximately ⅓ of patients. There are two types ofactivating mutations in FLT3 described in patients with leukemia. Theseinclude a spectrum of internal tandem duplications (ITD) occurringwithin the auto-inhibitory juxtamembrane domain (Nakao M, et al.,Leukemia, 1996, 10:1911-1918; Thiede C, et al., Blood, 2002,99:4326-4335), and activation loop mutations that include Asp835Tyr(D835Y), Asp835Val (D835V), Asp835.1His (D835H), Asp835Glu (D835E),Asp835Ala (D835A), Asp835Asn (D835N), Asp835 deletion and Ile836deletion (Yamamoto Y₁ et al., Blood 2001, :97:2434-2439; Abu-Duhier F M,et al., Br. J. Haematol., 2001, 113:983-988). Internal tandemduplication (ITD) mutations within the JM domain contribute to about17-34% of FLT3 activating mutations in AML. FLT3-ITD has also beendetected at low frequency in myelodysplastic syndrome (MDS) (Yokota S,et al., Leukemia, 1997, 11:1605-4609; Horiike S. et al., Leukemia, 1997,11:1442-1446). The ITDs are always in-frame, and are limited to the JMdomain. However, they vary in length and position from patient topatient. These repeat sequences may serve to disrupt the autoinhibitoryactivity of the JM domain resulting in the constitutive activation ofFLT3. Both FLT3-ITD and FLT3-Asp835 mutations are associated with FLT3autophosphoryiation and phosphorylation of downstream targets (Mizuki M,et al. Blood, 2000, 96:3907-3914; Mizuki M, et al. Blood, 2003,101:3164-3173; Hayakawa F, et al., Oncogene, 2000, 19: 624-631).

Inhibitors of FLT3 are presently being studied and have reached clinicaltrials as monotherapy in relapsed or refractory AML patients, some orall of whom had FLT3 mutations. Collectively, these data suggest thatFLT3 is an attractive therapeutic target for the development of kinaseinhibitors for AML and other associated diseases.

Janus kinase (JAK) is a family of intracellular, non-receptor tyrosinekinases that transduce cytokine-mediated signals via the JAK-STATpathway. The JAK family plays a role in the cytokine-dependentregulation of proliferation and function of cells involved in immuneresponse. Cytokines bind to their receptors, causing receptordimerization, and this enables JAKs to phosphorylate each other as wellas specific tyrosine motifs within the cytokine receptors. STATs thatrecognize these phosphotyrosine motifs are recruited to the receptor,and are then themselves activated by a JAK-dependent tyrosinephosphorylation event. Upon activation, STATs dissociate from thereceptors, dimerize, and translocate to the nucleus to bind to specificDNA sites and alter transcription.

Currently, there are four known mammalian JAK family members: JAK1(Janus kinase-1), JAK2 (Janus kinase-2), JAK3 (Janus kinase, leukocyte;JAKL; L-JAK and Janus kinase-3) and TYK2 (protein-tyrosine kinase 2).While JAK1, JAK2 and TYK2 are ubiquitously expressed, JAK3 is reportedto be preferentially expressed in natural killer (NK) cells and notresting T cells (“Biology and significance of the JAK/STAT signalingpathways.” Growth Factors, April 2012; 30(2): 88).

JAK1 is essential for signaling for certain type I and type II cytokinesIt interacts with the common gamma chain (γc) of type I cytokinereceptors to elicit signals from the IL-2 receptor family, the IL-4receptor family, the gp130 receptor family. It is also important fortransducing a signal by type I (IFN-α/β) and type II (IFN-γ)interferons, and members of the IL-10 family via type II cytokinereceptors. Genetic and biochemical studies have shown that JAK1 isfunctionally and physically associated with the type I interferon (e g.,IFNalpha), type II interferon (e.g., IFNgamma), IL-2 and IL-6 cytokinereceptor complexes. Furthermore, characterization of tissues derivedfrom JAK1 knockout mice demonstrated critical roles for this kinase inthe IFN, IL-IO, IL-2/IL-4, and IL-6 pathways.

Expression of JAK1 in cancer cells enables individual cells to contract,potentially allowing them to escape their tumor and metastasize to otherparts of the body. Elevated levels of cytokines which signal throughJAK1 have been implicated in a number of immune and inflammatorydiseases. JAK1 or JAK family kinase inhibitors may be useful formodulating or treating in such diseases. (Kisseleva et al., 2002, Gene285:1-24; Levy et al., 2005, Nat. Rev. Mol. Cell Biol. 3:651-662). Ahumanized monoclonal antibody targeting the IL-6 pathway (Tocilizumab)was approved by the European Commission for the treatment ofmoderate-to-severe rheumatoid arthritis (Scheinecker et al., 2009, Nat.Rev. Drug Discov. 8:273-274).

JAK2 is implicated in signaling by members of the type II cytokinereceptor family (e.g. interferon receptors), the GM-CSF receptor family,the gp130 receptor family. JAK2 signaling is activated downstream fromthe prolactin receptor. Studies have identified a high prevalence of anacquired activating JAK2 mutation (JAK2V617F) in myleoproliferativedisorders such as polycythemia vera, essential thrombocythemia andidiopathic myelofibrosis, etc. The mutant JAK2 protein is able toactivate downstream signaling in the absence of cytokine stimulation,resulting in autonomous growth and/or hypersensitivity to cytokines andis believed to play a role in driving these diseases. Additionalmutations or translocations resulting dysregulated JAK2 function havebeen described in other malignancies (Ihle J. N. and Gilliland D. G.,Curr. Opin. Genet. Dev., 2007, 17:8; Sayyah J. and Sayeski P. P., Curr.Oncol. Rep., 2009, 11: 117). Inhibitors of JAK2 have been described tobe useful in myeloproliferative diseases (Santos et al, Blood, 2010,115:1131; Barosi G. and Rosti V., Curr. Opin. Hematol, 2009, 16:129,Atallah E. and Versotvsek S., Exp. Rev. Anticancer Ther. 2009, 9:663).

JAK3 associates exclusively with the gamma common cytokine receptorchain, which is present in the IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21cytokine receptor complexes. JAK3 is predominantly expressed in immunecells and transduces a signal in response to its activation via tyrosinephosphorylation by interleukin receptors. Since JAK3 expression isrestricted mostly to hematopoietic cells, its role in cytokine signalingis thought to be more restricted than other JAKs. Mutations of JAK3result in severe combined immunodeficiency (SCID). (O'Shea et al., 2002,Cell, 109 (suppl.): S121-S131). Based on its role in regulatinglymphocytes, JAK3 and JAK3-mediated pathways have been targeted forimmunosuppressive indications (e.g., transplantation rejection andrheumatoid arthritis) (Baslund et al., 2005, Arthritis & Rheumatism52:2686-2692; Changelian et al., 2003, Science 302: 875-878).

TYK2 is implicated in IFN-α, IL-6, IL-10 and IL-12 signaling.Biochemical studies and gene-targeted mice uncovered the crucial role ofTYK2 in immunity. Tyk2-deficient mice are viable and fertile but displaymultiple immunological defects, most prominently high sensitivity toinfections and defective tumor surveillance. In contrast, inhibition ofTYK2 results in increased resistance against allergic, autoimmune andinflammatory diseases. Particularly, targeting Tyk2 appears to be apromising strategy for the treatment of IL-12-, IL-23- or Type 1IFN-mediated diseases. These include but are not limited to rheumatoidarthritis, multiple sclerosis, lupus, psoriasis, psoriatic arthritis,inflammatory bowel disease, uveitis, sarcoidosis, and tumors (Shaw, M.et al, Proc. Natl. Acad. Sci. USA, 2003, 100, 11594-11599; Ortmann, R.A., and Shevach, E. M. Clin. Immunol, 2001, 98, 109-118; Watford et al,Immunol. Rev., 2004, 202: 139). [“Janus Kinase (JAK) Inhibitors inRheumatoid Arthritis.” Current Rheumatology Reviews, 2011, 7, 306-312].

A fully human monoclonal antibody targeting the shared p40 subunit ofthe IL-12 and 11-23 cytokines (Ustekinumab) was recently approved by theEuropean Commission for the treatment of moderate-to-severe plaquepsoriasis (Krueger et al., 2007, N. Engl. J. Med. 356:580-92; Reich etal., 2009, Nat. Rev. Drug Discov. 8:355-356). In addition, an antibodytargeting the IL-12 and IL-23 pathways underwent clinical trials fortreating Crohn's Disease (Mannon et al., 2004, N.Engl. J. Med. 351:2069-79).

When dysregulated, JAK-mediated responses can positively or negativelyaffect cells leading to over-activation and malignancy or immune andhematopoietic deficiencies, respectively, and suggests the utility foruse of inhibitors of JAK kinases. The JAK/STAT signaling pathway isinvolved in a variety of hyperproliferative and cancer-related processesincluding cell-cycle progression, apoptosis, angiogenesis, invasion,metastasis and evasion of the immune system (Haura et al., NatureClinical Practice Oncology, 2005, 2(6), 315-324; Verna et al., Cancerand Metastasis Reviews, 2003, 22, 423-434). In addition, the JAK/STATsignaling pathway is important in the genesis and differentiation ofhematopoietic cells and regulating both pro- and anti-inflammatory andimmune responses (O'Sullivan et al., Molecular Immunology 2007,44:2497).

Therefore, the JAK/STAT pathway, and in particular all four members ofthe JAK family, are believed to play a role in the pathogenesis of theasthmatic response, chronic obstructive pulmonary disease, bronchitis,and other related inflammatory diseases of the lower respiratory tract.The JAK/STAT pathway has also been implicated to play a role ininflammatory diseases/conditions of the eye including, but not limitedto, iritis, uveitis, scleritis, conjunctivitis, as well as chronicallergic responses. Since cytokines utilize different patterns of JAKkinases (O'Sullivan et al., Mol. Immunol, 2007, 44:2497; Murray J.,Immunol, 2007, 178:2623), there may be utility for antagonists of JAKkinases with differing intra-family selectivity profiles in diseasesassociated with particular cytokines or in diseases associated withmutations or polymorphisms in the JAK/STAT pathways.

Rheumatoid arthritis (RA) is an autoimmune disease characterized bychronic joint inflammation. Patients with rheumatoid arthritis treatedwith JAK inhibitor showed that inhibition of JAK1 and JAK3 blockssignalling by multiple cytokines that are important for lymphocytefunction, including interleukin-2 (IL-2), IL-4, IL-7, IL-9, IL-15 andIL-21. [Fleischmann, R. et al., “Placebo-controlled trial of tofacitinibmonotherapy in rheumatoid arthritis.” N. Engl. J. Med., 367, 495-507(2012)]. It was conjectured that small-molecule inhibitors that directlyinactivate specific JAK isoforms would also reduce not only the clinicalsymptoms of RA, but also suppress the upregulation of many of theproinflammatory cytokines that are critical in driving RA diseaseprogression. [“Inhibitors of JAK for the treatment of rheumatoidarthritis: rationale and clinical data.” Clin. Invest., (2012) 2(1),39-47]

Persistent activation of STAT3 or STAT5 has been demonstrated in a widespectrum of solid human tumors including breast, pancreatic, prostate,ovarian and hepatic carcinomas, as well as in the majority ofhematopoietic tumors including lymphomas and leukemias. In this context,inactivation of JAK/STAT signaling in many hematopoietic tumors resultedin inhibition of cell proliferation and/or induction of apoptosis.Although STAT3 in tumor cells can be activated by various kinases, JAK2has been shown to be the most important upstream activator mediatingSTAT3 activation in human tumor cell lines derived from various solidtumors [Mohamad Bassam Sonbol, Belal Firwana, Ahmad Zarzour, MohammadMorad, Vishal Rana and Ramon V. Tiu “Comprehensive review of JAKinhibitors in myeloproliferative neoplasms.” Therapeutic Advances inHematology 2013, 4(1), 15-35; Hedvat M, Huszar D, Herrmann A, Gozgit JM, Schroeder A, Sheehy A, et al. “The JAK2 inhibitor AZD1480 potentlyblocks Stat3 signaling and oncogenesis in solid tumors.” Cancer Cell2009; 16(6):487-97.]. Therefore, inhibition of JAK kinases may have abeneficial role in the therapeutic treatment of these diseases.

Clearly, protein kinase inhibitors have gathered attention as a new drugcategory for both immunosuppresion and antiinflammatory drug, and forcancer drug. Thus, new or improved agents which inhibit protein kinasessuch as Aurora inhibitors, FLT3 inhibitors and Janus kinases inhibitorsare continually needed that act as immunosuppressive agents for organtransplants, and antitumor agents, as well as agents for the preventionand treatment of autoimmune diseases (e.g., multiple sclerosis,psoriasis, rheumatoid arthritis, asthma, type I diabetes, inflammatorybowel disease, Crohn's disease, polycythemia vera, essentialthrombocythemia, myelofibrosis, autoimmune thyroid disorders,Alzheimer's disease), diseases involving a hyperactive inflammatoryresponse (e.g., eczema), allergies, chronic obstructive pulmonarydisease, bronchitis, cancer (e.g., prostate, acute myelogenous leukemia,chronic myelogenous leukemia, acute lymphocytic leukemia, leukemia,multiple myeloma), and some immune reactions (e.g., skin rash or contactdermatitis or diarrhea) caused by other therapeutics, to name a few. Thecompounds, compositions and methods described herein are directed towardthese needs and other ends.

SUMMARY OF THE INVENTION

The invention provides compounds that inhibit, regulate, and/or modulateone or more protein kinases such as JAK, FLT3 and Aurora kinaseactivities, and are useful for treating proliferative diseases,autoimmune diseases, allergic diseases, inflammatory diseases,transplantation rejections, and their co-morbidities. This inventionalso provides methods of making the compound, methods of using suchcompounds in the treatment of said diseases in mammals, especially inhumans, and pharmaceutical compositions containing these compounds. Thecompounds or the pharmaceutical composition disclosed herein have betterprospects for clinical application. Compared with the similar compounds,the compounds disclosed herein have a better pharmacological activity,pharmacokinetic properties, physical and chemical properties and/orlesser toxicity. In particular, the compounds of the present inventiondisplay potent inhibitory activities against target kinases, andoptimized selectivity and exhibit good absorption and highbioavailability in vivo pharmacokinetic experiments. In addition, thecompounds or the pharmaceutical composition disclosed herein have goodmembrane permeability and solubility.

Specifically, in one aspect, provided herein is a compound havingFormula (I):

or a stereoisomer, a tautomer, an N-oxide, a solvate, a metabolite, apharmaceutically acceptable salt or a prodrug thereof, wherein each ofZ, Z¹, A and R¹ is as defined herein.

In one embodiment, Z is C₇-C₁₂ spiro bicycloalkyl, C₇-C₁₂ fusedbicycloalkyl, 7-12 membered spiro heterobicyclyl or 7-12 membered fusedheterobicycloalkyl, wherein Z is optionally substituted by 1, 2, 3, 4 or5 R² groups;

Z¹ is H, C₁-C₁₂ alkyl, C₃-C₁₂ cycloalkyl or 3-12 membered heterocyclyl,wherein Z¹ is optionally substituted by 1, 2, 3, 4 or 5 R³ groups;

A is indazolyl or pyrazolopyridinyl, wherein A is optionally substitutedby 1, 2, 3, 4 or 5R⁴ groups;

R¹ is H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl,C₁-C₁₂ alkoxyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, 3-12membered heterocyclyl, C₆-C₁₂ aryl, 5-12 membered heteroaryl,—(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein R¹ is optionally substituted by 1, 2, 3, 4 or 5R⁸ groups;

each R² is independently H, F, Cl, Br, I, NO₂, N₃, CN, OH, NH₂,—C(═O)CH₂CN, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl, C₁-C₁₂ alkoxy, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl, 5-12 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—C(═NR^(c))NR^(a)R^(b), —N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵or —S(═O)₂NR^(a)R^(b), or two adjacent R² taken together with the atomsto which they are attached form a C₃-C₁₂ cycloalkyl or 3-12 memberedheterocycloalkyl group, wherein each of the above substituents isoptionally independently substituted by 1, 2, 3, 4 or 5R⁸ groups;

each R³ and R⁴ is independently H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, —(C₁-C₄alkylene)-(C₃-C₁₂ cycloalkyl), C₆-C₁₂ aryl, 3-12 membered heterocyclyl,—(C₁-C₄ alkylene)-(3-12 membered heterocyclyl), 5-12 memberedheteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein each R³ and R⁴ is optionally independently substituted by 1, 2,3, 4 or 5R⁸ groups;

each R⁵ is independently H, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl or 5-12 membered heteroaryl, wherein each R⁵ is optionallyindependently substituted by 1, 2, 3, 4 or 5R⁸ groups;

each R⁶ and R⁷ is independently H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl,3-12 membered heterocyclyl or 5-12 membered heteroaryl, or R⁶ and R⁷taken together with the carbon atom to which they are attached form aC₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 membered heterocyclyl or 5-12membered heteroaryl group, wherein each of the above substituents isoptionally independently substituted by 1, 2, 3, 4 or 5R⁸ groups;

each R⁸ is independently F, Cl, Br, I, CN, NO₂, N₃, C₁-C₁₂ alkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl, 5-12 membered heteroaryl, NH₂, —NH(C₁-C₁₂ alkyl),—NH(CH₂)_(n)—(C₃-C₁₂ cycloalkyl), —NH(CH₂)_(n)—(C₆-C₁₂ aryl),—NH(CH₂)_(n)— (3-12 membered heterocyclyl), —NH(CH₂)_(n)-(5-12 memberedheteroaryl), —N(C₁-C₁₂ alkyl)₂, —N[(CH₂)_(n)—(C₃-C₁₂ cycloalkyl)]₂,—N[(CH₂)_(n)—(C₆-C₁₂ aryl)]₂, —N[(CH₂)_(n)-(3-12 memberedheterocyclyl)]₂, —N[(CH₂)_(n)-(5-12 membered heteroaryl)]₂, OH,—O(C₁-C₁₂ alkyl), —O(CH₂)_(n)—(C₃-C₁₂cycloalkyl),—O(CH₂)_(n)—(C₆-C₁₂aryl), —O(CH₂)_(n)-(3-12 membered heterocyclyl) or—O(CH₂)_(n)-(5-12 membered heteroaryl);

each R^(a), R^(b) and R^(c) is independently H, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₄ alkylene)-(C₃-C₆cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₄ alkylene)-(3-6 memberedheterocyclyl), C₆-C₁₀ aryl, —(C₁-C₄ alkylene)-(C₆-C₁₀ aryl), 5-10membered heteroaryl or —(C₁-C₄ alkylene)-(5-10 membered heteroaryl), orR^(a) and R^(b) taken together with the nitrogen atom to which they areattached form a 3-8 membered heterocyclyl group, wherein each of theabove substituents is optionally independently substituted by 1, 2, 3 or4 substitutents independently selected from F, Cl, Br, CN, N₃, OH, NH₂,C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy and C₁-C₆ alkylamino;

each m is independently 1 or 2; and

each n is independently 0, 1, 2, 3 or 4.

In another embodiment, Z is C₈-C₁₁ spiro bicycloalkyl, C₈-C₁₀ fusedbicycloalkyl, 8-11 membered spiro heterobicyclyl or 8-10 membered fusedheterobicycloalkyl, wherein Z is optionally substituted by 1, 2, 3 or4R² groups.

In one embodiment, Z¹ is H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl or 3-6membered heterocyclyl, wherein Z¹ is optionally substituted by 1, 2 or3R³ groups.

In another embodiment, R¹ is H, F, Cl, CN, N₃, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxyl, C₃-C₆cycloalkyl, 3-6 membered heterocyclyl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or—S(═O)₂NR^(a)R^(b), wherein R¹ is optionally substituted by 1, 2 or 3R⁸groups.

In one embodiment, each R² is independently H, F, Cl, CN, N₃, NO₂, OH,NH₂, —C(═O)CH₂CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆haloalkyl, C₁-C₆ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)—R⁵,—O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),or two adjacent R² taken together with the atoms to which they areattached form a C₃-C₆ cycloalkyl, or 3-6 membered heterocycloalkylgroup, wherein each of the above substituents is optionallyindependently substituted by 1, 2 or 3R⁸ groups.

In another embodiment, each R³ and R⁴ is independently H, F, Cl, Br, I,NO₂, N₃, CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, —(C₁-C₂ alkylene)-(C₃-C₆ cycloalkyl), phenyl, 3-6 memberedheterocyclyl, —(C₁-C₂ alkylene)-(3-6 membered heterocyclyl), 5-6membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b),—C(═O)R⁵, —OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b), wherein each R³ and R⁴ isoptionally independently substituted by 1, 2 or 3R⁸ groups.

In one embodiment, each R⁵ is independently H, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6membered heterocyclyl or 5-6 membered heteroaryl, wherein each R⁵ isoptionally independently substituted by 1, 2 or 3R⁸ groups.

In another embodiment, each R⁶ and R⁷ is independently H, F, Cl, Br, I,CN, N₃, NO₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, phenyl, 3-6 membered heterocyclyl or 5-6 memberedheteroaryl, or R⁶ and R⁷ taken together with the carbon atom to whichthey are attached form a C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl or 5-6 membered heteroaryl group, wherein each of the abovesubstituents is optionally independently substituted by 1, 2 or 3R⁸groups.

In one embodiment, each R⁸ is independently F, Cl, CN, N₃, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, NH₂, —NH(C₁-C₆ alkyl),—NH(CH₂)_(n)—(C₃-C₆ cycloalkyl), —NH(CH₂)_(n)-phenyl, —NH(CH₂)_(n)-(3-6membered heterocyclyl), —NH(CH₂)_(n)-(5-6 membered heteroaryl), —N(C₁-C₄alkyl)₂, —N[(CH₂)_(n)—(C₃-C₆cycloalkyl)]₂, —N[(CH₂)_(n)-phenyl]₂,—N[(CH₂)_(n)-(3-6 membered heterocyclyl)]₂, —N[(CH₂)_(n)-(5-6 memberedheteroaryl)]₂, OH, —O(C₁-C₆ alkyl), —O(CH₂)_(n)—(C₃-C₆ cycloalkyl),—O(CH₂)_(n)-phenyl, —O(CH₂)_(n)-(3-6 membered heterocyclyl) or—O(CH₂)_(n)-(5-6 membered heteroaryl).

In another embodiment, each R^(a), R^(b) and R^(c) is independently H,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₂alkylene)-(C₃-C₆ cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₂alkylene)-(3-6 membered heterocyclyl), phenyl, —(C₁-C₂ alkylene)-phenyl,5-6 membered heteroaryl or —(C₁-C₂ alkylene)-(5-6 membered heteroaryl),or R^(a) and R^(b) taken together with the nitrogen atom to which theyare attached form a 3-6 membered heterocyclyl group, wherein each of theabove substituents is optionally independently substituted by 1, 2 or 3substitutents independently selected from F, Cl, CN, N₃, OH, NH₂, C₁-C₄alkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy and C₁-C₄ alkylamino.

In one embodiment, Z is:

or a stereoisomer thereof, wherein each X, X′, X² and X³ isindependently CH₂, NH or O, with the proviso that when X² is O, X³ isnot O; Z is optionally substituted by 1, 2 or 3R² groups.

In another embodiment, A is:

wherein A is optionally substituted by 1, 2 or 3R⁴ groups.

In one embodiment, Z¹ is H, methyl, ethyl, n-propyl, isopropyl orcyclopropyl.

In another embodiment, R¹ is H, F, Cl, CN, N₃, C₁-C₄ alkyl, C₂-C₄alkenyl, C₂-C₄ alkynyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxyl, C₃-C₆cycloalkyl, 3-6 membered heterocyclyl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or—S(═O)₂NR^(a)R^(b), wherein R¹ is optionally substituted by 1, 2 or 3R⁸groups.

In one embodiment, each R² is independently H, F, Cl, CN, N₃, NO₂, OH,NH₂, —C(═O)CH₂CN, C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₁-C₄haloalkyl, C₁-C₄ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)—R⁵,—O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b), wherein each R² isoptionally independently substituted by 1, 2 or 3R⁸ groups.

In still another embodiment, each R⁵ is independently H, C₁-C₄ alkyl,C₁-C₄ haloalkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, phenyl,3-6 membered heterocyclyl or 5-6 membered heteroaryl, wherein each R⁵ isoptionally independently substituted by 1, 2 or 3R⁸ groups.

In another aspect, provided herein is a pharmaceutical compositioncomprising the compound disclosed herein, and a pharmaceuticallyacceptable excipient, carrier, adjuvant, vehicle or a combinationthereof.

In one embodiment, the pharmaceutical composition disclosed hereinfurther comprising a therapeutic agent selected from the groupconsisting of chemotherapeutic agents, anti-proliferative agents,phosphodiesterase 4 (PDE4) inhibitors, β₂-adrenoreceptor agonists,corticosteroids, non-steroidal GR agonists, anticholinergic agents,antihistamine, anti-inflammatory agents, immunosuppressants,immunomodulators, agents for treating atherosclerosis, agents fortreating pulmonary fibrosis and combinations thereof.

In another aspect, provided herein is a method of preventing, treatingor lessening the severity of a protein kinase-mediated disease in apatient by administering to the patient with the compound disclosedherein or the pharmaceutical composition disclosed herein.

In one embodiment, the protein kinase-mediated disease is JAK-, FLT3- orAurora-mediated disease.

In another embodiment, the protein kinase-mediated disease is aproliferative disease, an autoimmune disease, an allergic disease, aninflammatory disease or a transplantation rejection.

In another embodiment, the protein kinase-mediated disease is cancer,polycythemia vera, essential thrombocytosis, myelofibrosis, chronicmyelogenous leukemia (CML), acute myeloid leukemia (AML), acutelymphocytic leukemia (ALL), chronic obstruction pulmonary disease(COPD), asthma, systemic lupus erythematosis, cutaneous lupuserythematosis, lupus nephritis, dermatomyositis, Sjogren's syndrome,psoriasis, type I diabetes mellitus, allergic airway disease, sinusitis,eczema, hives, food allergies, allergies to insect venom, inflammatorybowel syndrome, Chron's disease, rheumatoid arthritis, juvenilearthritis, psoriatic arthritis, organ transplant rejection, tissuetransplant rejection or cell transplant rejection.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in preventing, treating orlessening the severity of a protein kinase-mediated disease in apatient.

In another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for treating cancer, polycythemia vera, essentialthrombocytosis, myelofibrosis, chronic myelogenous leukemia (CML), acutemyeloid leukemia (AML), acute lymphocytic leukemia (ALL), chronicobstruction pulmonary disease (COPD), asthma, systemic and cutaneouslupus erythematosis, lupus nephritis, dermatomyositis, Sjogren'ssyndrome, psoriasis, type I diabetes mellitus, allergic airway disease,sinusitis, eczema, hives, food allergies, allergies to insect venom,inflammatory bowel syndrome, Crohn's disease, rheumatoid arthritis,juvenile arthritis, psoriatic arthritis, organ transplant rejection,tissue transplant rejection or cell transplant rejection.

In another aspect, provided herein is a method of modulating theactivity of a protein kinase with the compound or the pharmaceuticalcomposition disclosed herein.

In one embodiment, the protein kinase is JAK kinase, FLT3 kinase, Aurorakinase or a combination thereof.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in modulating the activity of aprotein kinase.

In still another aspect, provided herein is use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for modulating the activity of a protein kinase.

In another aspect, provided herein are methods for preparation,separation and purification of the compounds represented by Formula (I).

Biological test results indicate that the compounds provided herein canbe used as preferable inhibitors of protein kinases.

Any embodiment disclosed herein can be combined with other embodimentsas long as they are not contradictory to one another, even though theembodiments are described under different aspects of the invention. Inaddition, any technical feature in one embodiment can be applied to thecorresponding technical feature in other embodiment as long as they arenot contradictory to one another, even though the embodiments aredescribed under different aspects of the invention.

The foregoing merely summarizes certain aspects of the invention and isnot intended to be limiting in nature. These aspects and other aspectsand embodiments are described more fully below.

DETAILED DESCRIPTION OF THE INVENTION Definitions and GeneralTerminology

Reference will now be made in detail to certain embodiments of theinvention, examples of which are illustrated in the accompanyingstructures and formulas. The invention is intended to cover allalternatives, modifications, and equivalents which may be includedwithin the scope of the present invention as defined by the claims. Oneskilled in the art will recognize many methods and materials similar orequivalent to those described herein, which could be used in thepractice of the present invention. The present invention is in no waylimited to the methods and materials described herein. In the event thatone or more of the incorporated literature, patents, and similarmaterials differs from or contradicts this application, including butnot limited to defined terms, term usage, described techniques, or thelike, this application controls.

It is further appreciated that certain features of the invention, whichare, for clarity, described in the context of separate embodiments, canalso be provided in combination in a single embodiment. Conversely,various features of the invention which are, for brevity, described inthe context of a single embodiment, can also be provided separately orin any suitable subcombination.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one skilled in theart to which this invention belongs. All patents and publicationsreferred to herein are incorporated by reference in their entirety.

As used herein, the following definitions shall apply unless otherwiseindicated. For purposes of this invention, the chemical elements areidentified in accordance with the Periodic Table of the Elements, CASversion, and the Handbook of Chemistry and Physics, 75^(th) Ed. 1994.Additionally, general principles of organic chemistry are described inSorrell et al., “Organic Chemistry”, University Science Books,Sausalito: 1999, and Smith et al., “March's Advanced Organic Chemistry”,John Wiley & Sons, New York: 2007, all of which are incorporated byreference in their entireties.

The grammatical articles “a”, “an” and “the”, as used herein, areintended to include “at least one” or “one or more” unless otherwiseindicated herein or clearly contradicted by the context. Thus, thearticles are used herein to refer to one or more than one (i.e. at leastone) of the grammatical objects of the article. By way of example, “acomponent” means one or more components, and thus, possibly, more thanone component is contemplated and may be employed or used in animplementation of the described embodiments.

As used herein, the term “subject” refers to an animal. Typically theanimal is a mammal. A subject also refers to for example, primates(e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats,rabbits, rats, mice, fish, birds and the like. In certain embodiments,the subject is a primate. In yet other embodiments, the subject is ahuman.

As used herein, “patient” refers to a human (including adults andchildren) or other animal. In one embodiment, “patient” refers to ahuman.

The term “comprising” is meant to be open ended, including the indicatedcomponent but not excluding other elements.

“Stereoisomers” refers to compounds which have identical chemicalconstitution, but differ with regard to the arrangement of the atoms orgroups in space. Stereoisomers include enantiomer, diastereomers,conformer (rotamer), geometric (cis/trans) isomer, atropisomer, etc.

“Chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g. melting points,boiling points, spectral properties or biological activities. Mixture ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography such as HPLC.

Stereochemical definitions and conventions used herein generally followParker et al., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York and Eliel et al., “Stereochemistry ofOrganic Compounds”, John Wiley & Sons, Inc., New York, 1994.

Many organic compounds exist in optically active forms, i.e., they havethe ability to rotate the plane of plane-polarized light. In describingan optically active compound, the prefixes D and L, or R and S, are usedto denote the absolute configuration of the molecule about its chiralcenter(s). The prefixes d and l or (+) and (−) are employed to designatethe sign of rotation of plane-polarized light by the compound, with (−)or l meaning that the compound is levorotatory. A compound prefixed with(+) or d is dextrorotatory. A specific stereoisomer may be referred toas an enantiomer, and a mixture of such stereoisomers is called anenantiomeric mixture. A 50:50 mixture of enantiomers is referred to as aracemic mixture or a racemate, which may occur where there has been nostereoselection or stereospecificity in a chemical reaction or process.

Any asymmetric atom (e.g., carbon or the like) of the compound(s)disclosed herein can be present in racemic or enantiomerically enriched,for example the (R)-, (S)- or (R,S)-configuration. In certainembodiments, each asymmetric atom has at least 50% enantiomeric excess,at least 60% enantiomeric excess, at least 70% enantiomeric excess, atleast 80% enantiomeric excess, at least 90% enantiomeric excess, atleast 95% enantiomeric excess, or at least 99% enantiomeric excess inthe (R)- or (S)-configuration.

Depending on the choice of the starting materials and procedures, thecompounds can be present in the form of one of the possiblestereoisomers or as mixtures thereof, such as racemates anddiastereoisomer mixtures, depending on the number of asymmetric carbonatoms. Optically active (R)- and (S)-isomers may be prepared usingchiral synthons or chiral reagents, or resolved using conventionaltechniques. If the compound contains a double bond, the substituent maybe E or Z configuration. If the compound contains a disubstitutedcycloalkyl, the cycloalkyl substituent may have a cis- ortrans-configuration.

Any resulting mixtures of stereoisomers can be separated on the basis ofthe physicochemical differences of the constituents, into the pure orsubstantially pure geometric isomers, enantiomers, diastereomers, forexample, by chromatography and/or fractional crystallization.

Any resulting racemates of final products or intermediates can beresolved into the optical antipodes by methods known to those skilled inthe art, e.g., by separation of the diastereomeric salts thereof.Racemic products can also be resolved by chiral chromatography, e.g.,high performance liquid chromatography (HPLC) using a chiral adsorbent.Preferred enantiomers can also be prepared by asymmetric syntheses. See,for example, Jacques, et al., Enantiomers, Racemates and Resolutions(Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis(2^(nd) Ed. Robert et al., Elsevier, Oxford, UK, 2012); Eliel et al.,Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilenet al., Tables of Resolving Agents and Optical Resolutions p. 268 (E. L.Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind., 1972). ChiralSeparation Techniques: A Practical Approach (Subramanian, G. Ed.,Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

The term “tautomer” or “tautomeric form” refers to structural isomers ofdifferent energies which are interconvertible via a low energy barrier.Where tautomerization is possible (e.g. in solution), a chemicalequilibrium of tautomers can be reached. For example, proton tautomers(also known as prototropic tautomers) include interconversions viamigration of a proton, such as keto-enol and imine-enamineisomerizations. Valence tautomers include interconversions byreorganization of some of the bonding electrons. A specific example ofketo-enol tautomerization is the interconversion of pentane-2,4-dioneand 4-hydroxypent-3-en-2-one tautomers. Another example oftautomerization is phenol-keto tautomerization. A specific example ofphenol-keto tautomerization is the interconversion of pyridin-4-ol andpyridin-4(1H)-one tautomers. Unless otherwise stated, all tautomericforms of the compounds disclosed herein are within the scope of theinvention.

As described herein, compounds disclosed herein may optionally besubstituted with one or more substituents, such as those illustratedbelow, or as exemplified by particular classes, subclasses, and speciesof the invention. It will be appreciated that the phrase “optionallysubstituted” is used interchangeably with the phrase “substituted orunsubstituted”. The term “optional” or “optionally” means that thesubsequently described event or circumstance may but need not occur, andthat the description includes instances where the event or circumstanceoccurs and instances in which it does not. In general, the term“substituted” refers to the replacement of one or more hydrogen radicalsin a given structure with the radical of a specified substituent. Unlessotherwise indicated, an optionally substituted group may have asubstituent at each substitutable position of the group. When more thanone position in a given structure can be substituted with more than onesubstituent selected from a specified group, the substituent may beeither the same or different at each position.

Some non-limiting examples of the substituents include D, F, Cl, Br, I,CN, N₃, —CN, —NO₂, —OH, —SH, —NH₂, —C(═O)CH₂CN, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(b)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—C(═NR^(c))NR^(a)R^(b), —N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵,—C(═O)NR^(a)R^(b), alkyl, haloalkyl, alkenyl, alkynyl, alkoxyl,alkylthiolyl, alkylamino, cycloalkyl, heterocyclyl, aryl and heteroaryl,and the like, wherein each R⁵, R⁶, R⁷, R^(a), R^(b), R^(c), m and ncarry the definitions described herein.

At various places in the present specification, substituents ofcompounds disclosed herein are disclosed in groups or in ranges. It isspecifically intended that the invention include each and everyindividual subcombination of the members of such groups and ranges. Forexample, the term “C₁-C₆ alkyl” is specifically intended to individuallydisclose methyl, ethyl, C₃ alkyl, C₄ alkyl, C₅ alkyl, and C₆ alkyl.

At various places in the present specification, linking substituents aredescribed. Where the structure clearly requires a linking group, theMarkush variables listed for that group are understood to be linkinggroups. For example, if the structure requires a linking group and theMarkush group definition for that variable lists “alkyl” or “aryl” thenit is understood that the “alkyl” or “aryl” represents a linkingalkylene group or arylene group, respectively.

The term “alkyl” or “alkyl group” refers to a saturated linear orbranched-chain monovalent hydrocarbon radical of 1 to 20 carbon atoms,wherein the alkyl radical may be optionally substituted independentlywith one or more substituents described below. Unless otherwisespecified, the alkyl group contains 1-20 carbon atoms. In oneembodiment, the alkyl group contains 1-12 carbon atoms. In anotherembodiment, the alkyl group contains 1-6 carbon atoms. In still anotherembodiment, the alkyl group contains 1-4 carbon atoms. In yet anotherembodiment, the alkyl group contains 1-3 carbon atoms. The alkyl radicalmay be optionally substituted independently with one or moresubstituents described herein.

Some non-limiting examples of the alkyl group include methyl (Me, —CH₃),ethyl (Et, —CH₂CH₃), 1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl(i-Pr, i-propyl, —CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃),2-methyl-1-propyl (i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl,—CH(CH₃)CH₂CH₃), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl(n-pentyl, —CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, 1-heptyl, 1-octyl, and the like.

The term “alkylene” refers to a saturated divalent hydrocarbon groupderived from a straight or branched chain saturated hydrocarbon by theremoval of two hydrogen atoms. Unless otherwise specified, the alkylenegroup contains 1-12 carbon atoms. In one embodiment, the alkylene groupcontains 1-6 carbon atoms. In another embodiment, the alkylene groupcontains 1-4 carbon atoms. In still another embodiment, the alkylenegroup contains 1-3 carbon atoms. In yet another embodiment, the alkylenegroup contains 1-2 carbon atoms. The alkylene group is exemplified bymethylene (—CH₂—), ethylidene (—CH₂CH₂—), isopropylidene (—CH(CH₃)CH₂—),and the like.

The term “alkenyl” refers to a linear or branched-chain monovalenthydrocarbon radical of 2 to 12 carbon atoms with at least one site ofunsaturation, i.e., a carbon-carbon, sp² double bond, wherein thealkenyl radical may be optionally substituted independently with one ormore substituents described herein, and includes radicals having “cis”and “trans” orientations, or alternatively, “E” and “Z” orientations. Inone embodiment, the alkenyl group contains 2-8 carbon atoms. In anotherembodiment, the alkenyl group contains 2-6 carbon atoms. In stillanother embodiment, the alkenyl group contains 2-4 carbon atoms. Somenon-limiting examples of the alkenyl group include ethylenyl or vinyl(—CH═CH₂), allyl (—CH₂CH═CH₂), and the like. The alkenyl radical may beoptionally substituted independently with one or more substituentsdescribed herein.

The term “alkynyl” refers to a linear or branched-chain monovalenthydrocarbon radical of 2 to 12 carbon atoms with at least one site ofunsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynylradical may be optionally substituted independently with one or moresubstituents described herein. In one embodiment, the alkynyl groupcontains 2-8 carbon atoms. In another embodiment, the alkynyl groupcontains 2-6 carbon atoms. In still another embodiment, the alkynylgroup contains 2-4 carbon atoms. Some non-limiting examples of thealkynyl group include ethynyl (—C≡CH), propargyl (—CH₂C≡CH), propynyl(—C≡C—CH₃), and the like.

The term “alkoxy” refers to an alkyl group, as previously defined,attached to the principal carbon atom through an oxygen atom. Unlessotherwise specified, the alkoxy group contains 1-12 carbon atoms. In oneembodiment, the alkoxy group contains 1-6 carbon atoms. In anotherembodiment, the alkoxy group contains 1-4 carbon atoms. In still anotherembodiment, the alkoxy group contains 1-3 carbon atoms. The alkoxyradical may be optionally substituted independently with one or moresubstituents described herein.

Some non-limiting examples of alkoxy groups include methoxy (MeO,—OCH₃), ethoxy (EtO, —OCH₂CH₃), 1-propoxy (n-PrO, n-propoxy,—OCH₂CH₂CH₃), 2-propoxy (i-PrO, i-propoxy, —OCH(CH₃)₂), 1-butoxy (n-BuO,n-butoxy, —OCH₂CH₂CH₂CH₃), 2-methyl-1-propoxy (i-BuO, i-butoxy,—OCH₂CH(CH₃)₂), 2-butoxy (s-BuO, s-butoxy, —OCH(CH3)CH₂CH₃),2-methyl-2-propoxy (t-BuO, t-butoxy, —OC(CH₃)₃), 1-pentoxy (n-pentoxy,—OCH₂CH₂CH₂CH₂CH₃), 2-pentoxy (—OCH(CH₃)CH₂CH₂CH₃), 3-pentoxy(—OCH(CH₂CH₃)₂), 2-methyl-2-butoxy (—OC(CH₃)₂CH₂CH₃), 3-methyl-2-butoxy(—OCH(CH₃)CH(CH₃)₂), 3-methyl-1-butoxy (—OCH₂CH₂CH(CH₃)₂),2-methyl-1-butoxy (—OCH₂CH(CH₃)CH₂CH₃), and the like.

The term “haloalkyl”, “haloalkenyl” or “haloalkoxy” refers to alkyl,alkenyl, or alkoxy, as the case may be, substituted with one or morehalogen atoms. Some non-limiting examples of haloalkyl and haloalkoxyare include trifluoromethyl (—CF₃), trifluoromethoxy (—OCF₃) and thelike.

The term “carbocycle”, “carbocyclyl” or “carbocyclic ring” refers to amonovalent or multivalent non-aromatic, saturated or partiallyunsaturated ring having 3 to 12 carbon atoms as a monocyclic, bicyclicor tricyclic ring system. The carbobicyclyl refers to a spirocarbobicyclyl, a fused carbobicyclyl or a bridged carbobicyclyl. Somenon-limiting examples of carbocyclyl groups include cycloalkyl,cycloalkenyl, and cycloalkynyl. Further non-limiting examples ofcarbocyclyl group include cyclopropyl, cyclobutyl, cyclopentyl,1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl,cyclohexadienyl, and the like.

The term “cycloalkyl” refers to a monovalent or multivalent saturatedring having 3 to 12 carbon atoms as a monocyclic, bicyclic, or tricyclicring system. In one embodiment, the cycloalkyl contains 3-12 carbonatoms. In another embodiment, the cycloalkyl contains 3-8 carbon atoms.In another embodiment, the cycloalkyl contains 3-6 carbon atoms. Instill another embodiment, cycloalkyl may be a C₇-C₁₂ bicycloalkyl whichrefers to C₇-C₁₂ spiro, C₇-C₁₂ fused bicycloalkyl and C₇-C₁₂ bridgedbicycloalkyl. In yet another embodiment, cycloalkyl may be a C₈-C₁₁bicycloalkyl which refers to C₈-C₁₁ spiro, C₈-C₁₁ fused bicycloalkyl andC₈-C₁₁ bridged bicycloalkyl. The cycloalkyl radical may be optionallysubstituted independently with one or more substituents describedherein.

The term “heterocycle”, “heterocyclyl”, or “heterocyclic ring” as usedinterchangeably herein refers to a monovalent or multivalent, saturatedor partially unsaturated, non-aromatic monocyclic, bicyclic or tricyclicring containing 3-12 ring atoms of which at least one ring atom isselected from nitrogen, sulfur and oxygen, and which may, unlessotherwise specified, be carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. The heterocyclyl containssaturated heterocyclyl (i.e. heterocycloalkyl) and partially unsaturatedheterocyclyl. Some non-limiting examples of heterocyclyl includeoxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl,3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl,1,3-dioxolanyl, dithiolanyl, tetrahydropyranyl, dihydropyranyl,2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, dioxanyl, thioxanyl, dithianyl,homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl (e.g.1,4-oxazepinyl, 1,2-oxazepinyl), diazepinyl (e.g. 1,4-diazepinyl,1,2-diazepinyl), dioxpinyl (e.g. 1,4-dioxpinyl, 1,2-dioxpinyl),thiazepinyl (e.g. 1,4-thiazepinyl, 1,2-thiazepinyl),2-oxa-5-azabicyclo[2.2.1]hept-5-yl, 2-azaspiro[4.4]nonanyl,1,6-dioxaspiro[4.4]nonanyl, 2-azaspiro[4.5]decanyl,8-azaspiro[4.5]decanyl, 7-az aspiro[4.5]decanyl,3-azaspiro[5.5]undecanyl , 2-azaspiro[5.5]undecanyl,octahydro-1H-isoindolyl, octahydrocyclopenta[c]pyrrolyl, indolinyl,1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxolyl,hexahydrofuro[3,2-b]furanyl, decahydroisoquinolinyl, and the like. Somenon-limiting examples of heterocyclyl wherein —CH₂— group is replaced by—C(═O)— moiety are 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl,2-piperidinonyl and 3,5-dioxopiperidinyl. Some non-limiting examples ofheterocyclyl wherein the ring sulfur atom is oxidized are sulfolanyl,1,1-dioxotetrahydrothiophenyl, 1,1-dioxothiomorpholinyl,1,1-dioxotetrahydro-2H-thiopyranyl. The heterocyclyl group may beoptionally substituted with one or more substituents described herein.

In one embodiment, heterocyclyl may be a 3-8 membered heterocyclyl,which refers to a monovalent or multivalent, saturated or partiallyunsaturated, monocyclic ring containing 3-8 ring atoms, of which atleast one ring atom is selected from nitrogen, sulfur and oxygen, and ofwhich may, unless otherwise specified, be carbon or nitrogen linked, andof which a —CH₂— group can optionally be replaced by a —C(═O)— group.Ring sulfur atoms may be optionally oxidized to form S-oxides. Ringnitrogen atoms maybe optionally oxidized to form N-oxides. Somenon-limiting examples of 3-8 membered heterocyclyl include azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl,pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl,1,3-dioxolanyl, dithiolanyl, tetrahydropyranyl, dihydropyranyl,2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, dioxanyl, thioxanyl, dithianyl,homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,diazepinyl, thiazepinyl, and the like. Some non-limiting examples ofheterocyclyl wherein —CH₂— group is replaced by —C(═O)— moiety are2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinonyl and3,5-dioxopiperidinyl. Some non-limiting examples of heterocyclyl whereinthe ring sulfur atom is oxidized are sulfolanyl,1,1-dioxo-thiomorpholinyl, and the like. The 3-8 membered heterocyclylgroup may be optionally substituted with one or more substituentsdescribed herein.

In another embodiment, heterocyclyl may be a 3-6 membered heterocyclyl,which refers to a monovalent or multivalent, saturated or partiallyunsaturated, non-aromatic monocyclic ring containing 3-6 ring atoms, ofwhich at least one ring atom is selected from nitrogen, sulfur andoxygen, and of which may, unless otherwise specified, be carbon ornitrogen linked, and of which a —CH₂— group can optionally be replacedby a —C(═O)— group. Ring sulfur atoms may be optionally oxidized to formS-oxides. Ring nitrogen atoms maybe optionally oxidized to formN-oxides. Some non-limiting examples of 3-6 membered heterocyclylinclude azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl,3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl,1,3-dioxolanyl, dithiolanyl, tetrahydropyranyl, dihydropyranyl,2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, dioxanyl, thioxanyl and dithianyl. The 3-6membered heterocyclyl group may be optionally substituted with one ormore substituents described herein.

In another embodiment, heterocyclyl refers to a 7-12 memberedheterocyclyl, which refers to a monovalent or multivalent, saturated orpartially unsaturated spiro, fused or bridged heterobicyclyl ringcontaining 7-12 ring atoms, of which at least one ring atom is selectedfrom nitrogen, sulfur and oxygen, and which may, unless otherwisespecified, be carbon or nitrogen linked, and of which a —CH₂— group canoptionally be replaced by a —C(═O)— group. Ring sulfur atoms may beoptionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. Some non-limiting examples of 7-12membered heterocyclyl include indolinyl,1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxolyl,2-oxa-5-azabicyclo[2.2.1]hept-5-yl, 2-azaspiro[4.4]nonanyl (e.g.2-azaspiro[4.4]nonane-4-yl, 2-azaspiro[4.4]nonane-2-yl),1,6-dioxaspiro[4.4]nonanyl (e.g. 1,6-dioxaspiro[4.4]nonan-9-yl,1,6-dioxaspiro[4.4]nonane-4-yl), 2-azaspiro[4.5]decanyl (e.g.2-azaspiro[4.5]decane-8-yl, 2-azaspiro[4.5]decane-2-yl),7-azaspiro[4.5]decanyl (e.g. 7-azaspiro[4.5]decane-8-yl,7-azaspiro[4.5]decane-2-yl) 3-azaspiro[5.5]undecanyl (e.g.3-azaspiro[5.5]undecane-3-yl, 3-azaspiro[5.5]undecane-9-yl),2-azaspiro[5.5]undecanyl, 8-azaspiro[4.5]decanyl,decahydroisoquinolinyl, octahydro-1H-isoindolyl (e.g.octahydro-1H-isoindole-5-yl, octahydro-1H-isoindole-7-yl),octahydrocyclopenta[c]pyrrolyl (e.g. octahydrocyclopenta[c]pyrrole-5-yl,octahydrocyclopenta[c]pyrrole-2-yl), hexahydrofuro[3,2-b]furanyl (e.g.hexahydrofuro[3,2-b]furan-2-yl, hexahydrofuro[3,2-b]furan-3-yl), and thelike. The 7-12 membered heterocyclyl group may be optionally substitutedwith one or more substituents described herein.

In still one embodiment, heterocyclyl refers to a 7-12 membered spiroheterobicyclyl, which refers to a monovalent or multivalent, saturatedor partially unsaturated, non-aromatic, spiro heterobicyclyl ringcontaining 7-12 ring atoms, of which at least one ring atom is selectedfrom nitrogen, sulfur and oxygen, and which may, unless otherwisespecified, be carbon or nitrogen linked, and of which a —CH₂— group canoptionally be replaced by a —C(═O)— group. Ring sulfur atoms may beoptionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. The 7-12 membered spiroheterobicyclyl contains 7-12 membered saturated spiro heterobicyclyl(i.e. 7-12 membered spiro heterobicycloalkyl) and 7-12 memberedpartially unsaturated spiro heterobicyclyl. Some non-limiting examplesof 7-12 membered spiro heterobicyclyl include 2-azaspiro[4.4]nonanyl(e.g. 2-azaspiro[4.4]nonane-4-yl, 2-azaspiro[4.4]nonane-2-yl), 1,6-dioxaspiro[4.4]nonanyl (e.g. 1,6-dioxaspiro[4.4]nonan-9-yl,1,6-dioxaspiro[4.4]nonane-4-yl), 2-azaspiro[4.5]decanyl (e.g.2-azaspiro[4.5]decane-8-yl, 2-azaspiro[4.5]decane-2-yl),7-azaspiro[4.5]decanyl (e.g. 7-azaspiro[4.5]decane-8-yl,7-azaspiro[4.5]decane-2-yl), 3-azaspiro[5.5]undecanyl (e.g.3-azaspiro[5.5]undecane-3-yl, 3-azaspiro[5.5]undecane-9-yl),2-azaspiro[5.5]undecanyl, 8-azaspiro[4.5]decanyl, and the like. The 7-12membered spiro heterobicyclyl group may be optionally substituted withone or more substituents described herein.

In still another embodiment, heterocyclyl refers to a 8-11 memberedspiro heterobicyclyl, which refers to a amonovalent or multivalent,saturated or partially unsaturated, non-aromatic, spiro heterobicyclylring containing 8-11 ring atoms, of which at least one ring atom isselected from nitrogen, sulfur and oxygen, and which may, unlessotherwise specified, be carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. The 8-11 membered spiroheterobicyclyl contains 8-11 membered saturated spiro heterobicyclyl(i.e. 8-11 membered spiro heterobicycloalkyl) and 8-11 memberedpartially unsaturated spiro heterobicyclyl. Some non-limiting examplesof 8-11 membered spiro heterobicyclyl include 2-azaspiro[4.4]nonanyl(e.g. 2-azaspiro[4.4]nonane-4-yl, 2-azaspiro[4.4]nonane-2-yl),1,6-dioxaspiro[4.4]nonanyl (e.g. 1,6-dioxaspiro[4.4]nonan-9-yl,1,6-dioxaspiro[4.4]nonane-4-yl), 2-azaspiro[4.5]decanyl (e.g.2-azaspiro[4.5]decane-8-yl, 2-azaspiro[4.5]decane-2-yl),7-azaspiro[4.5]decanyl (e.g. 7-azaspiro[4.5]decane-8-yl,7-azaspiro[4.5]decane-2-yl), 3-azaspiro[5.5]undecanyl (e.g.3-azaspiro[5.5]undecane-3-yl, 3-azaspiro[5.5]undecane-9-yl),2-azaspiro[5.5]undecanyl , 8-azaspiro[4.5]decanyl, and the like. The8-11 membered Spiro heterobicyclyl group may be optionally substitutedwith one or more substituents described herein.

In yet another embodiment, heterocyclyl refers to a 7-12 membered fusedheterobicyclyl, which refers to a amonovalent or multivalent, saturatedor partially unsaturated, non-aromatic fused heterobicyclyl ringcontaining 7-12 ring atoms, of which at least one ring atom is selectedfrom nitrogen, sulfur and oxygen, and which may, unless otherwisespecified, be carbon or nitrogen linked, and of which a —CH₂— group canoptionally be replaced by a —C(═O)— group. Ring sulfur atoms may beoptionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. The 7-12 membered fusedheterobicyclyl contains 7-12 membered saturated fused heterobicyclyl(i.e. 7-12 membered fused heterobicycloalkyl) and 7-12 memberedpartially unsaturated fused heterobicyclyl. Some non-limiting examplesof 7-12 membered fused heterobicyclyl includeoctahydrocyclopenta[c]pyrrolyl, octahydro-1H-isoindolyl, indolinyl,1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxolyl,hexahydrofuro[3,2-b]furanyl, hexahydrofuro[2,3-b]furanyl,decahydroisoquinolinyl, and the like. The 7-12 membered fusedheterobicyclyl group may be optionally substituted with one or moresubstituents described herein.

The terms “fused bicyclic ring”, “fused cyclic”, “fused bicyclyl” and“fused cyclyl” are used interchangeably refer to a monovalent ormultivalent saturated or partially unsaturated, but not aromaticbicyclic ring system, and such that two rings share one common bond.Such a system may contain isolated or conjugated unsaturation, but notaromatic or heteroaromatic rings in its core structure (but may havearomatic substitution thereon).

The terms “spirocyclyl”, “spirocyclic”, “spiro bicyclyl” and “spirobicyclic” are used interchangeably and refer to a monovalent ormultivalent, saturated or partially unsaturated, ring system wherein aring originating from a particular annular carbon of another ring. Forexample, as depicted below in Structure a, a saturated ring system (ringB and B′) is termed as “fused bicyclyl”, whereas ring A and ring B sharean atom between the two saturated ring system, which terms as a“spirocyclyl” or “spiro bicyclyl”. Each ring in the fused bicyclyl orthe spiro bicyclyl can be either a carbocyclyl or a heterocyclyl, andeach ring is optionally substituted independently with one or moresubstituents described herein.

The term “heterocycloalkyl” refers to a monovalent or multivalentsaturated ring having 3 to 12 ring atoms as a monocyclic, bicyclic, ortricyclic ring system in which at least one ring atom is selected fromnitrogen, sulfur and oxygen and which may, unless otherwise specified,be carbon or nitrogen linked, and of which a —CH₂— group can optionallybe replaced by a —C(═O)— group. Ring sulfur atoms may be optionallyoxidized to form S-oxides. Ring nitrogen atoms maybe optionally oxidizedto form N-oxides. Some non-limiting examples of heterocycloalkyl includeazetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl,imidazolidinyl, tetrahydrothienyl, tetrahydrofuranyl, piperidinyl,piperazinyl, morpholinyl, dioxanyl, dithianyl, dithiolanyl,isoxazolidinyl, isothiazolidinyl, 1,2-oxazinanyl, 1,2-thiazinanyl,hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, oxepanyl,thiepanyl, oxazepinyl (e.g. 1,4-oxazepinyl, 1,2-oxazepinyl), diazepinyl(e.g. 1,4-diazepinyl, 1,2-diazepinyl), dioxpinyl (e.g. 1,4-dioxpinyl,1,2-dioxpinyl), thiazepinyl (e.g. 1,4-thiazepinyl, 1,2-thiazepinyl),2-azaspiro[4.4]nonanyl , 1,6-dioxaspiro[4.4]nonanyl,2-azaspiro[4.5]decanyl, 8-azaspiro[4.5]decanyl, 7-azaspiro[4.5]decanyl,3-azaspiro[5.5]undecanyl, 2-azaspiro[5.5]undecanyl,2-octahydro-1H-isoindolyl, octahydrocyclopenta[c]pyrrolyl,hexahydrofuro[3,2-b]furanyl, decahydroisoquinolinyl,hexahydrofuro[2,3-b]furanyl, and the like. The heterocycloalkyl groupmay be optionally substituted with one or more substituents describedherein.

In one embodiment, heterocycloalkyl refers to a 7-12 memberedheterocycloalkyl, which refers to a monovalent or multivalent saturatedspiro, fused or bridged heterobicycloalkyl, containing 7-12 ring atoms,of which at least one ring atom is selected from nitrogen, sulfur andoxygen and which may, unless otherwise specified, be carbon or nitrogenlinked, and of which a —CH₂— group can optionally be replaced by a—C(═O)— group. Ring sulfur atoms may be optionally oxidized to formS-oxides. Ring nitrogen atoms maybe optionally oxidized to formN-oxides. The 7-12 membered heterocycloalkyl group may be optionallysubstituted with one or more substituents described herein.

In one embodiment, heterocycloalkyl refers to a 3-6 memberedheterocycloalkyl, which refers to a monovalent or multivalent saturatedheterocyclyl ring containing 3-6 ring atoms, of which at least one ringatom is selected from nitrogen, sulfur and oxygen and which may, unlessotherwise specified, be carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. Some non-limiting examples of 3-6membered heterocycloalkyl include azetidinyl, oxetanyl, thietanyl,pyrrolidinyl, pyrazolidinyl, imidazolidinyl, piperidinyl, piperazinyl,morpholinyl, dioxanyl, dithianyl, dithiolanyl, isoxazolidinyl,isothiazolidinyl, and hexahydropyridazinyl. The 3-6 memberedheterocycloalkyl group may be optionally substituted with one or moresubstituents described herein.

In another embodiment, heterocycloalkyl refers to a 7-12 membered spiroheterobicycloalkyl, which refers to a monovalent or multivalentsaturated spiro heterobicycloalkyl ring containing 7-12 ring atoms, ofwhich at least one ring atom is selected from nitrogen, sulfur andoxygen. Unless otherwise specified, the 7-12 membered spiroheterobicycloalkyl maybe carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. Some non-limiting examples of 7-12membered spiro heterobicycloalkyl include 2-azaspiro[4.4]nonanyl,1,6-dioxaspiro[4.4]nonanyl, 2-azaspiro[4.5]decanyl,8-azaspiro[4.5]decanyl, 7-azaspiro[4.5]decanyl,3-azaspiro[5.5]undecanyl, 2-azaspiro[5.5]undecanyl, and the like. The7-12 membered spiro heterobicycloalkyl group may be optionallysubstituted with one or more substituents described herein.

In another embodiment, heterocycloalkyl refers to a 7-12 membered fusedheterobicycloalkyl, which refers to a monovalent or multivalentsaturated fused heterobicycloalkyl ring containing 7-12 ring atoms, ofwhich at least one ring atom is selected from nitrogen, sulfur andoxygen. Unless otherwise specified, the 7-12 membered fusedheterobicycloalkyl maybe carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. Some non-limiting examples of 7-12membered fused heterobicycloalkyl include octahydro-1H-isoindolyl (e.g.octahydro-1H-isoindole-5-yl, octahydro-1H-isoindole-7-yl),octahydrocyclopenta[c]pyrrolyl (e.g. octahydrocyclopenta[c]pyrrole-5-yl,octahydrocyclopenta[c]pyrrole-2-yl), hexahydrofuro[3,2-b]furanyl (e.g.hexahydrofuro[3,2-b]furan-2-yl, hexahydrofuro[3,2-b]furan-3-yl),decahydroisoquinolinyl, hexahydrofuro[2,3-b]furanyl, and the like. The7-12 membered fused heterocybicloalkyl group may be optionallysubstituted with one or more substituents described herein.

In another embodiment, heterocycloalkyl refers to a 8-10 membered fusedheterobicycloalkyl, which refers to a monovalent or multivalentsaturated fused heterobicycloalkyl ring containing 8-10 ring atoms, ofwhich at least one ring atom is selected from nitrogen, sulfur andoxygen. Unless otherwise specified, the 8-10 membered fusedheterobicycloalkyl maybe carbon or nitrogen linked, and of which a —CH₂—group can optionally be replaced by a —C(═O)— group. Ring sulfur atomsmay be optionally oxidized to form S-oxides. Ring nitrogen atoms maybeoptionally oxidized to form N-oxides. Some non-limiting examples of 8-10membered fused heterobicycloalkyl include octahydro-1H-isoindolyl (e.g.octahydro-1H-isoindole-5-yl, octahydro-1H-isoindole-7-yl),octahydrocyclopenta[c]pyrrolyl (e.g. octahydrocyclopenta[c]pyrrole-5-yl,octahydrocyclopenta[c]pyrrole-2-yl), hexahydrofuro[3,2-b]furanyl (e.g.hexahydrofuro[3,2-b]furan-2-yl, hexahydrofuro[3,2-b]furan-3-yl),decahydroisoquinolinyl, hexahydrofuro[2,3-b]furanyl, and the like. The8-10 membered fused heterocybicloalkyl group may be optionallysubstituted with one or more substituents described herein.

The term “n membered” where n is an integer typically describes thenumber of ring-forming atoms in a moiety where the number ofring-forming atoms is n. For example, piperidinyl is an example of a 6membered heterocycloalkyl and 1,2,3,4-tetrahydronaphthalenyl is anexample of a 10 membered carbocyclyl group.

The term “unsaturated” refers to a moiety having one or more units ofunsaturation.

The term “heteroatom” refers to one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon, including any oxidized form of nitrogen, sulfur,or phosphorus; the quaternized form of any basic nitrogen; or asubstitutable nitrogen of a heterocyclic ring, for example N (as in3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR (as inN-substituted pyrrolidinyl).

The term “halogen” refers to Fluoro (F), Chloro (Cl), Bromo (Br), orIodo (I).

The term “azido” or “N₃” refers to an azide moiety. This radical may beattached, for example, to a methyl group to form azidomethane (methylazide, MeN₃); or attached to a phenyl group to form phenyl azide (PhN₃).

The term “aryl” refers to monocyclic, bicyclic, and tricycliccarbocyclic ring systems having a total of 6 to 14 ring members,preferably, 6 to 12 ring members, and more preferably 6 to 10 ringmembers, wherein at least one ring in the system is aromatic, whereineach ring in the system contains 3 to 7 ring members and that has one ormore points of attachment to the rest of the molecule. The term “aryl”may be used interchangeably with the term “aryl ring” or “aromaticring”. Some non-limiting examples of the aryl group would includephenyl, naphthyl, and anthracenyl. The aryl radical is optionallysubstituted independently with one or more substituents describedherein.

The term “heteroaryl” or “heteroaromatic ring” refers to monocyclic,bicyclic, and tricyclic ring systems having a total of 5 to 12 ringmembers, preferably, 5 to 10 ring members, and more preferably 5 to 6ring members, wherein at least one ring in the system is aromatic, atleast one aromatic ring in the system contains one or more heteroatoms,wherein each ring in the system contains 5 to 7 ring members and thathas one or more points of attachment to the rest of the molecule. Theterm “heteroaryl” may be used interchangeably with the term “heteroarylring” or the term “heteroaromatic ring”. In one embodiment, heteroarylrefers to a 5-12 membered heteroaryl comprises 1, 2, 3 or 4 heteroatomsindependently selected from O, S and N. In another embodiment,heteroaryl refers to a 5-10 membered heteroaryl comprises 1, 2, 3 or 4heteroatoms independently selected from O, S and N. In anotherembodiment, heteroaryl refers to a 5-6 membered heteroaryl comprises 1,2, 3 or 4 heteroatoms independently selected from O, S and N.Theheteroaryl radical is optionally independently substituted with one ormore substituents described herein.

Some non-limiting examples of the heteroaryl group include 2-furanyl,3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl,5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl,4-pyridyl, pyridonyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl,pyrimidonyl, pyrimidinedionyl, pyridazinyl (e.g., 3-pyridazinyl),2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl),triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl,pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl,1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl,1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl,1,3,5-triazinyl, and the following bicycles: benzimidazolyl, benzofuryl,benzothiophenyl, indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g.,2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g.,1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl),imidazo[1,2-c]pyridyl, pyrazolo[1,5-c]pyridyl, pyrazolo[4,3-c]pyridinyl,pyrazolo[3,4-b]pyridinyl, pyrazolo[1,5-c]pyrimidyl,imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl,[1,2,4]triazolo[1,5-a]pyrimidinyl and [1,2,4]triazolo[1,5-a]pyridyl.

The term “azolyl” refers to a 5-membered or 9-membered heteroaryl ringsystem containing at least one nitrogen atom. Some non-limiting examplesof the azolyl include pyrazolyl, imidazolyl, oxazolyl, isoxazolyl,oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, diazolyl, triazolyl,indazolyl, pyrazolo[3,4-b]pyridinyl,pyrazolo[4,3-c]pyridinyl,1H-imidazo[4,5-b]pyridinyl and1H-benzo[d]imidazolyl, and the like.

The term “carboxy” or “carboxyl”, whether used alone or with otherterms, such as “carboxyalkyl”, refers to —CO₂H. The term “carbonyl”,whether used alone or with other terms, such as “aminocarbonyl”, denotes—(C═O)—.

The term “alkylamino” embraces “N-alkylamino” and “N,N-dialkylamino”where amino groups are independently substituted with one alkyl radicalor with two alkyl radicals, respectively. In one embodiment alkylaminoare “lower alkylamino” radicals having one or two alkyl radicals of oneto six carbon atoms, attached to a nitrogen atom. In another embodimentalkylamino are alkylamino radicals having one or two alkyl radicals ofone to four carbon atoms, attached to a nitrogen atom. Some non-limitingexamples of alkylamino include N-methylamino, N-ethylamino,N,N-dimethylamino, N,N-diethylamino, and the like.

The term “arylamino” refers to amino groups, which have been substitutedwith one or two aryl radicals, such as N-phenylamino. The arylaminoradicals may be further substituted on the aryl ring portion of theradical.

The term “aminoalkyl” refers to linear or branched alkyl radicals havingone to about ten carbon atoms any one of which may be substituted withone or more amino radicals. More preferred aminoalkyl radicals are“lower aminoalkyl” radicals having 1-6 carbon atoms and one or moreamino radicals. Examples of such radicals include aminomethyl,aminoethyl, aminopropyl, aminobutyl and aminohexyl.

As described herein, a bond drawn from a substituent to the center ofone ring within a ring system (as shown below) represents substitutionof the substituent at any substitutable position on the ring system. Forexample, as depicted below, Figure b represents possible substitution inany of the positions on the ring C and ring D shown in Figurec˜Structure g.

As described herein, a connecting bond drawn from the center of one ringwithin a ring system (as shown in Structure h, wherein each X and X′ isindependently CH₂, NH or O) represents connection of the connecting bondattached to the rest of the molecule at any substitutable position onthe ring system. For example, Structure h represents possible connectionattached to the rest of the molecule in any of the position on ring Eand ring F.

As described herein, two connecting bonds drawn from the center of onering within a ring system (as shown in Structure i) representsconnection of the connecting bonds attached to the rest of the moleculeat any two substitutable positions on the ring system. For example,Structure i represents possible connection attached to the rest of themolecule in any two of the positions on ring G.

The term “protecting group” or “PG” refers to a substituent that iscommonly employed to block or protect a particular functionality whilereacting other functional groups on the compound. For example, an“amino-protecting group” is a substituent attached to an amino groupthat blocks or protects the amino functionality in the compound.Suitable amino-protecting groups include acetyl, trifluoroacetyl,t-butoxy-carbonyl (BOC, Boc), benzyloxycarbonyl (CBZ, Cbz) and9-fluorenylmethylenoxy-carbonyl (Fmoc). Similarly, a “hydroxy-protectinggroup” refers to a substituent of a hydroxy group that blocks orprotects the hydroxy functionality. Suitable protecting groups includeacetyl and silyl. A “carboxy-protecting group” refers to a substituentof the carboxy group that blocks or protects the carboxy functionality.Common carboxy-protecting groups include —CH₂CH₂SO₂Ph, cyanoethyl,2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxy-methyl,2-(p-toluenesulfonyl)-ethyl, 2-(p-nitrophenylsulfenyl)-ethyl,2-(diphenylphosphino)-ethyl, nitroethyl and the like. For a generaldescription of protecting groups and their use, see T. W. Greene,Protective Groups in Organic Synthesis, John Wiley & Sons, New York,1991; and P. J. Kocienski, Protecting Groups, Thieme, Stuttgart, 2005.

The term “prodrug” as used herein, represents a compound that istransformed in vivo into a compound of Formula (I). Such atransformation can be affected, for example, by hydrolysis in blood orenzymatic transformation of the prodrug form to the parent form in bloodor tissue. Prodrugs of the compounds disclosed herein may be, forexample, esters. Esters that may be utilized as prodrugs in the presentinvention are phenyl esters, aliphatic (C₁-C₂₄) esters, acyloxymethylesters, carbonates, carbamates, and amino acid esters. For example, acompound disclosed herein that contains an OH group may be acylated atthis position in its prodrug form. Other prodrug forms includephosphates, such as, for example those phosphates resulting from thephosphonation of an OH group on the parent compound. A thoroughdiscussion of prodrugs is provided in Higuchi et al., Pro-drugs as NovelDelivery Systems, Vol. 14, A.C.S. Symposium Series; Roche et al.,Bioreversible Carriers in Drug Design, American PharmaceuticalAssociation and Pergamon Press, 1987; Rautio et al., Prodrugs: Designand Clinical Applications, Nat. Rev. Drug Discovery, 2008, 7, 255-270,and Hecker et al., Prodrugs of Phosphates and Phosphonates, J. Med.Chem., 2008, 51, 2328-2345, all of which are incorporated herein byreference.

A “metabolite” refers to a product produced through metabolism in thebody of a specified compound or salt thereof. The metabolites of acompound may be identified using routine techniques known in the art andtheir activities determined using tests such as those described herein.Such products may result for example from the oxidation, reduction,hydrolysis, amidation, deamidation, esterification, deesterification,enzymatic cleavage, and the like, of the administered compound.Accordingly, the invention includes metabolites of compounds disclosedherein, including compounds produced by a process comprising contactinga compound disclosed herein with a mammal for a period of timesufficient to yield a metabolic product thereof.

A “pharmaceutically acceptable salt” refers to organic or inorganicsalts of a compound disclosed herein. The pharmaceutically acceptablesalts are well known in the art. For example, Berge et al., describepharmaceutically acceptable salts in detail in J. Pharm. Sci., 1977, 66,1-19, which is incorporated herein by reference. Some non-limitingexamples of the pharmaceutically acceptable salt include salts of anamino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, oxalic acid, maleic acid,tartaric acid, citric acid, succinic acid or malonic acid.

Other examples of the pharmaceutically acceptable salt include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike.

Pharmaceutically acceptable salts derived from appropriate bases includealkali metal, alkaline earth metal, ammonium and N⁺(C₁-C₄ alkyl)₄ salts.This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. Water oroil-soluble or dispersible products may be obtained by suchquaternization. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further examples of the pharmaceutically acceptable salt include, whenappropriate, nontoxic ammonium, quaternary ammonium, and amine cationsformed using counterions such as halide, hydroxide, carboxylate,sulfate, phosphate, nitrate, C₁-C₈ sulfonate and aryl sulfonate.

A “solvate” refers to an association or complex of one or more solventmolecules and a compound disclosed herein. Examples of solvents thatform solvates include, but are not limited to, water, isopropanol,ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.The term “hydrate” refers to the complex where the solvent molecule iswater.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers in one embodiment, to ameliorating thedisease or disorder (i.e., slowing or arresting or reducing thedevelopment of the disease or at least one of the clinical symptomsthereof). In another embodiment “treat”, “treating” or “treatment”refers to alleviating or ameliorating at least one physical parameterincluding those which may not be discernible by the patient. In yetanother embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to preventing ordelaying the onset or development or progression of the disease ordisorder.

“Inflammatory disorder/disease” as used herein can refer to any disease,disorder, or syndrome in which an excessive or unregulated inflammatoryresponse leads to excessive inflammatory symptoms, host tissue damage,or loss of tissue function. “Inflammatory disorder/disease” also refersto a pathological state mediated by influx of leukocytes and/orneutrophil chemotaxis.

“Inflammation” as used herein refers to a localized, protective responseelicited by injury or destruction of tissues, which serves to destroy,dilute, or wall off (i.e. sequester) both the injurious agent and theinjured tissue. Inflammation is notably associated with influx ofleukocytes and/or neutrophil chemotaxis. Inflammation can result frominfection with pathogenic organisms and viruses and from noninfectiousmeans such as trauma or reperfusion following myocardial infarction orstroke, immune response to foreign antigen, and autoimmune responses.Accordingly, inflammatory disorders amenable to treatment with thecompounds disclosed herein encompass disorders associated with reactionsof the specific defense system as well as with reactions of thenonspecific defense system.

“Specific defense system” refers to the component of the immune systemthat reacts to the presence of specific antigens. Examples ofinflammation resulting from a response of the specific defense systeminclude the classical response to foreign antigens, autoimmune diseases,and delayed type hypersensitivity response mediated by T-cells. Chronicinflammatory diseases, the rejection of solid transplanted tissue andorgans, e.g., kidney and bone marrow transplants, and graft versus hostdisease (GVHD), are further examples of inflammatory reactions of thespecific defense system.

“Autoimmune disease” as used herein refers to any group of disorders inwhich tissue injury is associated with humoral or cell-mediatedresponses to the body's own constituents.

“Allergic disease” as used herein refers to any symptoms, tissue damage,or loss of tissue function resulting from allergy. “Arthritic disease”as used herein refers to any disease that is characterized byinflammatory lesions of the joints attributable to a variety ofetiologies. “Dermatitis” as used herein refers to any of a large familyof diseases of the skin that are characterized by inflammation of theskin attributable to a variety of etiologies. “Transplant rejection” asused herein refers to any immune reaction directed against graftedtissue, such as organs or cells (e.g., bone marrow), characterized by aloss of function of the grafted and surrounding tissues, pain, swelling,leukocytosis, and thrombocytopenia. The therapeutic methods of thepresent invention include methods for the treatment of disordersassociated with inflammatory cell activation.

The terms “cancer” and “cancerous” refer to or describe thephysiological condition in mammals that is typically characterized byunregulated cell growth. A “tumor” comprises one or more cancerouscells. Examples of cancer include, but are not limited to, carcinoma,lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. Moreparticular examples of such cancers include squamous cell cancer (e.g.,epithelial squamous cell cancer), lung cancer including small-cell lungcancer, non-small cell lung cancer (“NSCLC”), adenocarcinoma of the lungand squamous carcinoma of the lung, cancer of the peritoneum,hepatocellular cancer, gastric or stomach cancer includinggastrointestinal cancer, pancreatic cancer, glioblastoma, cervicalcancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breastcancer, colon cancer, rectal cancer, colorectal cancer, endometrial oruterine carcinoma, salivary gland carcinoma, kidney or renal cancer,prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, analcarcinoma, penile carcinoma, as well as head and neck cancer.

Description of Compounds of the Invention

In the present invention, novel compounds which are inhibitors ofprotein kinase activity, in particular JAK kinase, FLT3 kinase andAurora kinase activity, are disclosed. Compounds which are proteinkinase inhibitors may be useful in the treatment of diseases associatedwith inappropriate protein kinase activity, in particular inappropriateJAK, FLT3 and Aurora kinase activity, for example in the treatment andprevention of diseases mediated by JAK kinase, FLT3 kinase and Aurorakinase involved signalling pathways. Such diseases include proliferativedisease, autoimmune disease, allergic disease, inflammatory disease,transplantation rejection, and their co-morbidities. In particular, acompound of the present invention may be useful in the treatment ofdiseases such as cancer, polycythemia vera, essential thrombocytosis,myelofibrosis, chronic myelogenous leukemia (CML), acute myeloidleukemia (AML), acute lymphocytic leukemia (ALL), chronic obstructionpulmonary disease (COPD), asthma, systemic and cutaneous lupuserythematosis, lupus nephritis, dermatomyositis, Sjogren's syndrome,psoriasis, type I diabetes mellitus, allergic airway disease, sinusitis,eczema, hives, food allergies, allergies to insect venom, inflammatorybowel syndrome, Crohn's disease, rheumatoid arthritis, juvenilearthritis, psoriatic arthritis, organ transplant rejection, tissuetransplant rejection, cell transplant rejection, to name a few.

In one embodiment, the compounds disclosed herein may show potentinhibitory activities against one or more protein kinases.

In one aspect, provided herein is a compound having Formula (I):

or a stereoisomer, a tautomer, an N-oxide, a solvate, a metabolite, apharmaceutically acceptable salt or a prodrug thereof, wherein each ofZ, Z¹, A and R¹ is as defined herein.

In one embodiment, Z is C₇-C₁₂ spiro bicycloalkyl, C₇-C₁₂ fusedbicycloalkyl, 7-12 membered spiro heterobicyclyl or 7-12 membered fusedheterobicycloalkyl, wherein Z is optionally substituted by 1, 2, 3, 4 or5R² groups;

Z¹ is H, C₁-C₁₂ alkyl, C₃-C₁₂ cycloalkyl or 3-12 membered heterocyclyl,wherein Z¹, except when Z¹ is H, is optionally substituted by 1, 2, 3, 4or 5R³ groups;

A is indazolyl or pyrazolopyridinyl, wherein A is optionally substitutedby 1, 2, 3, 4 or 5R⁴ groups;

R¹ is H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl,C₁-C₁₂ alkoxyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, 3-12membered heterocyclyl, C₆-C₁₂ aryl, 5-12 membered heteroaryl,—(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein when R¹ is not H, F, Cl, Br, I, NO₂, N₃ or CN, R¹ is optionallysubstituted by 1, 2, 3, 4 or 5R⁸ groups;

each R² is independently H, F, Cl, Br, I, NO₂, N₃, CN, OH, NH₂,—C(═O)CH₂CN, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl, C₁-C₁₂ alkoxy, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl, 5-12 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—C(═NR^(c))NR^(a)R^(b), —N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵or —S(═O)₂NR^(a)R^(b), or two adjacent R² taken together with the atomsto which they are attached form a C₃-C₁₂ cycloalkyl or 3-12 memberedheterocycloalkyl group, wherein each of the above substituents, exceptH, F, Cl, Br, I, NO₂, N₃ and CN, is optionally independently substitutedby 1, 2, 3, 4 or 5R⁸ groups;

each R³ and R⁴ is independently H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, —(C₁-C₄alkylene)-(C₃-C₁₂ cycloalkyl), C₆-C₁₂ aryl, 3-12 membered heterocyclyl,—(C₁-C₄ alkylene)-(3-12 membered heterocyclyl), 5-12 memberedheteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein when R³ or R⁴ is not H, F, Cl, Br, I, NO₂, N₃ or CN, R³ or R⁴respectively is optionally substituted by 1, 2, 3, 4 or 5R⁸ groups;

each R⁵ is independently H, C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl or 5-12 membered heteroaryl, wherein each R⁵, except whenR⁵ is H, is optionally independently substituted by 1, 2, 3, 4 or 5R⁸groups;

each R⁶ and R⁷ is independently H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl,3-12 membered heterocyclyl or 5-12 membered heteroaryl, or R⁶ and R⁷taken together with the carbon atom to which they are attached form aC₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 membered heterocyclyl or 5-12membered heteroaryl group, wherein each of the above substituents,except H, F, Cl, Br, I, NO₂, N₃ and CN, is optionally independentlysubstituted by 1, 2, 3, 4 or 5R⁸ groups;

each R⁸ is independently F, Cl, Br, I, CN, NO₂, N₃, C₁-C₁₂ alkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl, 5-12 membered heteroaryl, NH₂, —NH(C₁-C₁₂ alkyl),—NH(CH₂)_(n)—(C₃-C₁₂ cycloalkyl), —NH(CH₂)_(n)—(C₆-C₁₂ aryl),—NH(CH₂)_(n)-(3-12 membered heterocyclyl), —NH(CH₂)_(n)-(5-12 memberedheteroaryl), —N(C₁-C₁₂ alkyl)₂, —N[(CH₂)_(n)—(C₃-C₁₂ cycloalkyl)]₂,—N[(CH₂)_(n)—(C₆-C₁₂ aryl)]₂, —N[(CH₂)_(n)-(3-12 memberedheterocyclyl)]₂, —N[(CH₂)_(n)-(5-12 membered heteroaryl)]₂, OH,—O(C₁-C₁₂ alkyl), —O(CH₂)_(n)—(C₃-C₁₂cycloalkyl),—O(CH₂)_(n)—(C₆-C₁₂aryl), —O(CH₂)_(n)-(3-12 membered heterocyclyl) or—O(CH₂)_(n)-(5-12 membered heteroaryl);

each R^(a), R^(b) and R^(c) is independently H, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₄ alkylene)-(C₃-C₆cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₄ alkylene)-(3-6 memberedheterocyclyl), C₆-C₁₀ aryl, —(C₁-C₄ alkylene)-(C₆-C₁₀ aryl), 5-10membered heteroaryl or —(C₁-C₄ alkylene)-(5-10 membered heteroaryl), orR^(a) and R^(b) taken together with the nitrogen atom to which they areattached form a 3-8 membered heterocyclyl group, wherein each of theabove substituents, except H, is optionally independently substituted by1, 2, 3 or 4 substitutents independently selected from F, Cl, Br, CN,N₃, OH, NH₂, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy and C₁-C₆alkylamino;

each m is independently 1 or 2; and

each n is independently 0, 1, 2, 3 or 4.

In another embodiment, Z is C₈-C₁₁ spiro bicycloalkyl, C₈-C₁₀ fusedbicycloalkyl, 8-11 membered spiro heterobicyclyl or 8-10 membered fusedheterobicycloalkyl, wherein Z is optionally substituted by 1, 2, 3 or4R² groups.

In one embodiment, Z¹ is H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl or 3-6membered heterocyclyl, wherein Z¹, except when Z¹ is H, is optionallysubstituted by 1, 2 or 3R³ groups.

In another embodiment, R¹ is H, F, Cl, CN, N₃, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxyl, C₃-C₆cycloalkyl, 3-6 membered heterocyclyl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or—S(═O)₂NR^(a)R^(b), wherein when R¹ is not H, F, Cl, CN or N₃, R¹ isoptionally substituted by 1, 2 or 3R⁸ groups.

In one embodiment, each R² is independently H, F, Cl, CN, N₃, NO₂, OH,NH₂, —C(═O)CH₂CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆haloalkyl, C₁-C₆ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)—R⁵,—O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),or two adjacent R² taken together with the atoms to which they areattached form a C₃-C₆ cycloalkyl, or 3-6 membered heterocycloalkylgroup, wherein each of the above substituents, except H, F, Cl, CN, N₃and NO₂, is optionally independently substituted by 1, 2 or 3R⁸ groups.

In another embodiment, each R³ and R⁴ is independently H, F, Cl, Br, I,NO₂, N₃, CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, —(C₁-C₂ alkylene)-(C₃-C₆ cycloalkyl), phenyl, 3-6 memberedheterocyclyl, —(C₁-C₂ alkylene)-(3-6 membered heterocyclyl), 5-6membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b),—C(═O)R⁵, —OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b), wherein when R³ or R⁴ is notH, F, Cl, Br, I, NO₂, N₃ or CN, R³ or R⁴ respectively is optionallysubstituted by 1, 2 or 3R⁸ groups.

In one embodiment, each R⁵ is independently H, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6membered heterocyclyl or 5-6 membered heteroaryl, wherein each R⁵,except when R⁵ is H, is optionally independently substituted by 1, 2 or3R⁸ groups.

In another embodiment, each R⁶ and R⁷ is independently H, F, Cl, Br, I,CN, N₃, NO₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, phenyl, 3-6 membered heterocyclyl or 5-6 memberedheteroaryl, or R⁶ and R⁷ taken together with the carbon atom to whichthey are attached form a C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl or 5-6 membered heteroaryl group, wherein each of the abovesubstituents, except H, F, Cl, Br, I, CN, N3 and NO₂, is optionallyindependently substituted by 1, 2 or 3R⁸ groups.

In one embodiment, each R⁸ is independently F, Cl, CN, N₃, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, NH₂, —NH(C₁-C₆ alkyl),—NH(CH₂)_(n)—(C₃-C₆ cycloalkyl), —NH(CH₂)_(n)-phenyl, —NH(CH₂)_(n)-(3-6membered heterocyclyl), —NH(CH₂)_(n)-(5-6 membered heteroaryl), —N(C₁-C₄alkyl)₂, —N[(CH₂)_(n)—(C₃-C₆cycloalkyl)]₂, —N[(CH₂)_(n)-phenyl]₂,—N[(CH₂)_(n)-(3-6 membered heterocyclyl)]₂, —N[(CH₂)_(n)-(5-6 memberedheteroaryl)]₂, OH, —O(C₁-C₆ alkyl), —O(CH₂)_(n)—(C₃-C₆ cycloalkyl),—O(CH₂)_(n)-phenyl, —O(CH₂)_(n)-(3-6 membered heterocyclyl) or—O(CH₂)_(n)-(5-6 membered heteroaryl).

In another embodiment, each R^(a), R^(b) and R^(c) is independently H,C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₂alkylene)-(C₃-C₆ cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₂alkylene)-(3-6 membered heterocyclyl), phenyl, —(C₁-C₂ alkylene)-phenyl,5-6 membered heteroaryl or —(C₁-C₂ alkylene)-(5-6 membered heteroaryl),or R^(a) and R^(b) taken together with the nitrogen atom to which theyare attached form a 3-6 membered heterocyclyl group, wherein each of theabove substituents, except H, is optionally independently substituted by1, 2 or 3 substitutents independently selected from F, Cl, CN, N₃, OH,NH₂, C₁-C₄ alkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy and C₁-C₄ alkylamino.

In one embodiment, Z is:

or a stereoisomer thereof, wherein each X, X′, X² and X³ isindependently CH₂, NH or O, with the proviso that when X² is O, X³ isnot O; and wherein Z is optionally substituted by 1, 2 or 3 R² groups.

In another embodiment, A is:

wherein A is optionally substituted by 1, 2 or 3R⁴ groups.

In one embodiment, Z¹ is H, methyl, ethyl, n-propyl, isopropyl orcyclopropyl.

In another embodiment, R¹ is H, F, Cl, CN, N₃, C₁-C₄ alkyl, C₂-C₄alkenyl, C₂-C₄ alkynyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxyl, C₃-C₆cycloalkyl, 3-6 membered heterocyclyl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or—S(═O)₂NR^(a)R^(b), wherein R¹, except when R¹ is H, F, Cl, CN or N₃, isoptionally substituted by 1, 2 or 3R⁸ groups.

In one embodiment, each R² is independently H, F, Cl, CN, N₃, NO₂, OH,NH₂, —C(═O)CH₂CN, C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₁-C₄haloalkyl, C₁-C₄ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c),—(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)R⁵,—O(CR⁶R⁷)_(n)—OR^(c), —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b), wherein each R², except whenR² is H, F, Cl, CN, N3 or NO₂, is optionally independently substitutedby 1, 2 or 3R⁸ groups.

In still another embodiment, each R⁵ is independently H, C₁-C₄ alkyl,C₁-C₄ haloalkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, phenyl,3-6 membered heterocyclyl or 5-6 membered heteroaryl, wherein each R⁵,except when R⁵ is H, is optionally independently substituted by 1, 2 or3R⁸ groups.

In yet another embodiment, some non-limiting examples of the compounddisclosed herein, and their stereoisomer, tautomer, N-oxide, solvate,pharmaceutically acceptable salts and solvates thereof, are shown in thefollowing:

TABLE 1

(1)

(2)

(3)

(4)

(5)

(6)

(7)

(8)

(9)

(10)

(11)

(12) or

(13)

Unless otherwise stated, all stereoisomers, tautomers, solvates,metabolites, salts, and pharmaceutically acceptable prodrugs of thecompounds of Formula (I) are within the scope of the invention.

The compounds disclosed herein may contain asymmetric or chiral centers,and therefore exist in different stereoisomeric forms. It is intendedthat all stereoisomeric forms of compounds of Formula (I), including butnot limited to, diastereomers, enantiomers, atropisomers, conformers(rotamers) and geometric (cis/trans) isomers as well as mixtures thereofsuch as racemic mixtures, form part of the present invention.

In the structures shown herein, where the stereochemistry of anyparticular chiral atom is not specified, then all stereoisomers arecontemplated and included as the compounds of the invention. Wherestereochemistry is specified by a solid wedge or dashed linerepresenting a particular configuration, then that stereoisomer is sospecified and defined.

The compounds of Formula (I) may exist in different tautomeric forms,and all such forms are embraced within the scope of the invention, asdefined by the claims.

The compounds of Formula (I) can be in the form of salts. In oneembodiment, the salts are pharmaceutically acceptable salts. The phrase“pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith. In another embodiment, the salts are not necessarilypharmaceutically acceptable salts, and which may be useful asintermediates for preparing and/or purifying compounds of Formula (I)and/or for separating enantiomers of compounds of Formula (I).

Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids, e.g., acetate, aspartate, benzoate,besylate, bromide/hydrobromide, bicarbonate/carbonate,bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride,chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate,gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate,lactate, lactobionate, laurylsulfate, malate, maleate, malonate,mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate,nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate,propionate, stearate, succinate, subsalicylate, tartrate, tosylate andtrifluoroacetate salts.

Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example,acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,toluenesulfonic acid, sulfosalicylic acid, and the like.

Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases.

Inorganic bases from which salts can be derived include, for example,ammonium salts and metals from columns I to XII of the periodic table.In certain embodiments, the salts are derived from sodium, potassium,ammonium, calcium, magnesium, iron, silver, zinc, and copper;particularly suitable salts include ammonium, potassium, sodium, calciumand magnesium salts.

Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like. Certain organic amines includeisopropylamine, benzathine, cholinate, diethanolamine, diethylamine,lysine, meglumine, pip erazine and tromethamine.

The pharmaceutically acceptable salts of the present invention can besynthesized from a basic or acidic moiety, by conventional chemicalmethods. Generally, such salts can be prepared by reacting free acidforms of these compounds with a stoichiometric amount of the appropriatebase (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or thelike), or by reacting free base forms of these compounds with astoichiometric amount of the appropriate acid. Such reactions aretypically carried out in water or in an organic solvent, or in a mixtureof the two. Generally, use of non-aqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile is desirable, wherepracticable. Lists of additional suitable salts can be found, e.g., in“Remington's Pharmaceutical Sciences”, 20th ed., Mack PublishingCompany, Easton, Pa., (1985); and in “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH,Weinheim, Germany, 2002).

Furthermore, the compounds disclosed herein, including their salts, canalso be obtained in the form of their hydrates, or include othersolvents such as ethanol, DMSO, and the like, used for theircrystallization. The compounds of the present invention may inherentlyor by design form solvates with pharmaceutically acceptable solvents(including water); therefore, it is intended that the invention embraceboth solvated and unsolvated forms.

Any formula given herein is also intended to represent isotopicallyunenriched forms as well as isotopically enriched forms of thecompounds. Isotopically enriched compounds have structures depicted bythe formulas given herein except that one or more atoms are replaced byan atom having a selected atomic mass or mass number. Examples ofisotopes that can be incorporated into compounds of the inventioninclude isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous,fluorine, and chlorine, such as ²H (deuterium, D), ³H, ¹¹C, ¹³C, ¹⁴C,¹⁵N, ¹⁷O, ¹⁸O, ¹⁸F, ³¹P, ³²P, ³⁵S, ³⁶Cl, ¹²⁵I, respectively.

In another aspect, the compounds of the invention include isotopicallyenriched compounds as defined herein, for example those into whichradioactive isotopes, such as ³H, ¹⁴C and ¹⁸F, or those into whichnon-radioactive isotopes, such as ²H and ¹³C are present. Suchisotopically enriched compounds are useful in metabolic studies (with¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detectionor imaging techniques, such as positron emission tomography (PET) orsingle-photon emission computed tomography (SPECT) including drug orsubstrate tissue distribution assays, or in radioactive treatment ofpatients. In particular, an ¹⁸F-enriched compound may be particularlydesirable for PET or SPECT studies. Isotopically-enriched compounds ofFormula (I) can generally be prepared by conventional techniques knownto those skilled in the art or by processes analogous to those describedin the accompanying Examples and Preparations using an appropriateisotopically-labeled reagent in place of the non-labeled reagentpreviously employed.

Further, substitution with heavier isotopes, particularly deuterium(i.e., ²H or D) may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements or an improvement in therapeutic index. Itis understood that deuterium in this context is regarded as asubstituent of a compound of Formula (I). The concentration of such aheavier isotope, specifically deuterium, may be defined by the isotopicenrichment factor. The term “isotopic enrichment factor” as used hereinmeans the ratio between the isotopic abundance and the natural abundanceof a specified isotope. If a substituent in a compound of this inventionis denoted deuterium, such compound has an isotopic enrichment factorfor each designated deuterium atom of at least 3500 (52.5% deuteriumincorporation at each designated deuterium atom), at least 4000 (60%deuterium incorporation), at least 4500 (67.5% deuterium incorporation),at least 5000 (75% deuterium incorporation), at least 5500 (82.5%deuterium incorporation), at least 6000 (90% deuterium incorporation),at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97%deuterium incorporation), at least 6600 (99% deuterium incorporation),or at least 6633.3 (99.5% deuterium incorporation). Pharmaceuticallyacceptable solvates in accordance with the invention include thosewherein the solvent of crystallization may be isotopically substituted,e.g. D₂O, acetone-d₆, DMSO-d₆.

In another aspect, provided herein are intermediates for preparing thecompounds disclosed herein.

In another aspect, provided herein are methods of preparing, methods ofseparating, and methods of purifying the compounds disclosed herein.

In another aspect, provided herein is a pharmaceutical compositioncomprising a therapeutically effective amount of the compound disclosedherein, and a pharmaceutically acceptable excipient, carrier, adjuvant,vehicle or a combination thereof. In some embodiments, the compositionis a liquid, solid, semi-solid, gel, or an aerosol form.

In another aspect, provided herein is a method of treating a disease ordisorder modulated by one or more protein kinases such as JAK kinase,FLT3 kinase and Aurora kinase, comprising administering to a mammal inneed of such treatment an effective amount of a compound or apharmaceutical composition disclosed herein. In one embodiment, thedisease or disorder is selected from proliferative disease, autoimmunedisease, allergic disease, inflammatory disease or transplantationrejection.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment of disease ordisorder selected from proliferative disease, autoimmune disease,allergic disease, inflammatory disease or transplantation rejection.

In another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for the treatment of disease or disorder selected fromproliferative disease, autoimmune disease, allergic disease,inflammatory disease or transplantation rejection.

In still another aspect, provided herein is use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for modulating the activity of protein kinase.

Pharmaceutical Composition, Formulations and Administration of theCompounds of the Invention

The present invention provides a pharmaceutical composition that includea compound disclosed herein, or a compound listed in Table 1; and apharmaceutically acceptable excipient, carrier, adjuvant, vehicle or acombination thereof. The amount of compound in the pharmaceuticalcomposition disclosed herein is such that is effective to detectablyinhibit a protein kinase in a biological sample or in a patient.

It will also be appreciated that certain compounds disclosed herein canexist in free form for treatment, or where appropriate, as apharmaceutically acceptable derivative thereof. Some non-limitingexamples of pharmaceutically acceptable derivative includepharmaceutically acceptable prodrugs, salts, esters, salts of suchesters, or any other adduct or derivative which upon administration to apatient in need is capable of providing, directly or indirectly, acompound as otherwise described herein, or a metabolite or residuethereof.

The pharmaceutical compositions disclosed herein may be prepared andpackaged in bulk form wherein a safe and effective amount of thecompound disclosed herein can be extracted and then given to the patientsuch as with powders or syrups. Alternatively, the pharmaceuticalcompositions disclosed herein may be prepared and packaged in unitdosage form wherein each physically discrete unit contains the compounddisclosed herein. When prepared in unit dosage form, the pharmaceuticalcompositions of the invention typically may contain, for example, from0.5 mg to 1 g, or from 1 mg to 700 mg, or from 5 mg to 100 mg of thecompound disclosed herein.

As used herein, “pharmaceutically acceptable excipient” means apharmaceutically acceptable material, composition or vehicle involved ingiving form or consistency to the pharmaceutical composition. Eachexcipient must be compatible with the other ingredients of thepharmaceutical composition when commingled such that interactions whichwould substantially reduce the efficacy of the compound disclosed hereinwhen administered to a patient and interactions which would result inpharmaceutical compositions that are not pharmaceutically acceptable areavoided. In addition, each excipient must bepharmaceutically-acceptable, e.g., of sufficiently high purity.

Suitable pharmaceutically acceptable excipients will vary depending uponthe particular dosage form chosen. In addition, suitablepharmaceutically acceptable excipients may be chosen for a particularfunction that they may serve in the composition. For example, certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the production of uniform dosage forms. Certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the production of stable dosage forms. Certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the carrying or transporting of the compound or compoundsdisclosed herein once administered to the patient from one organ, orportion of the body, to another organ, or portion of the body. Certainpharmaceutically acceptable excipients may be chosen for their abilityto enhance patient compliance.

Suitable pharmaceutically acceptable excipients comprise the followingtypes of excipients: diluents, fillers, binders, disintegrants,lubricants, glidants, granulating agents, coating agents, wettingagents, solvents, co-solvents, suspending agents, emulsifiers,sweetners, flavoring agents, flavor masking agents, coloring agents,anticaking agents, hemectants, chelating agents, plasticizers, viscosityincreasing agents, antioxidants, preservatives, stabilizers,surfactants, and buffering agents. The skilled artisan will appreciatethat certain pharmaceutically acceptable excipients may serve more thanone function and may serve alternative functions depending on how muchof the excipient is present in the formulation and what other excipientsare present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enablethem to select suitable pharmaceutically-acceptable excipients inappropriate amounts for use in the invention. In addition, there are anumber of resources that are available to the skilled artisan whichdescribe pharmaceutically acceptable excipients and may be useful inselecting suitable pharmaceutically acceptable excipients. Examplesinclude Remington's Pharmaceutical Sciences (Mack Publishing Company),The Handbook of Pharmaceutical Additives (Gower Publishing Limited), andThe Handbook of Pharmaceutical Excipients (the American PharmaceuticalAssociation and the Pharmaceutical Press).

In Remington: The Science and Practice of Pharmacy, 21st edition, 2005,ed. D. B. Troy, Lippincott Williams & Wilkins, Philadelphia, andEncyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C.Boylan, 1988-1999, Marcel Dekker, New York, the contents of each ofwhich is incorporated by reference herein, are disclosed variouscarriers used in formulating pharmaceutically acceptable compositionsand known techniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds disclosedherein, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention.

The pharmaceutical compositions disclosed herein are prepared usingtechniques and methods known to those skilled in the art. Some of themethods commonly used in the art are described in Remington'sPharmaceutical Sciences (Mack Publishing Company).

Accordingly, in another aspect the invention is directed to process forthe preparation of a pharmaceutical composition comprising the compounddisclosed herein and a pharmaceutically acceptable excipient, carrier,adjuvant, vehicle or a combination thereof, which comprises mixing theingredients. A pharmaceutical composition comprising the compounddisclosed herein may be prepared by, for example, admixture at ambienttemperature and atmospheric pressure.

The compounds disclosed herein will typically be formulated into adosage form adapted for administration to the patient by the desiredroute of administration. For example, dosage forms include those adaptedfor (1) oral administration such as tablets, capsules, caplets, pills,troches, powders, syrups, elixers, suspensions, solutions, emulsions,granula, and cachets; (2) parenteral administration such as sterilesolutions, suspensions, and freeze drying powder; (3) transdermaladministration such as transdermal patches; (4) rectal administrationsuch as suppositories; (5) inhalation such as aerosols, solutions, anddry powders; and (6) topical administration such as creams, ointments,lotions, solutions, pastes, sprays, foams, and gels.

In one embodiment, the compounds disclosed herein will be formulated fororal administration. In another embodiment, the compounds disclosedherein will be formulated for inhaled administration. In a furtherembodiment, the compounds disclosed herein will be formulated forintranasal administration. In another embodiment, the compoundsdisclosed herein will be formulated for transdermal administration. In afurther embodiment, the compounds disclosed herein will be formulatedfor topical administration.

The pharmaceutical compositions provided herein can be provided ascompressed tablets, tablet triturates, chewable lozenges, rapidlydissolving tablets, multiple compressed tablets, or enteric-coatingtablets, sugar-coated, or film-coated tablets. Enteric-coated tabletsare compressed tablets coated with substances that resist the action ofstomach acid but dissolve or disintegrate in the intestine, thusprotecting the active ingredients from the acidic environment of thestomach. Enteric-coatings include, but are not limited to, fatty acids,fats, phenyl salicylate, waxes, shellac, ammoniated shellac, andcellulose acetate phthalates. Sugar-coated tablets are compressedtablets surrounded by a sugar coating, which may be beneficial incovering up objectionable tastes or odors and in protecting the tabletsfrom oxidation. Film-coated tablets are compressed tablets that arecovered with a thin layer or film of a water-soluble material. Filmcoatings include, but are not limited to, hydroxyethylcellulose, sodiumcarboxymethylcellulose, polyethylene glycol 4000, and cellulose acetatephthalate. Film coating imparts the same general characteristics assugar coating. Multiple compressed tablets are compressed tablets madeby more than one compression cycle, including layered tablets, andpress-coated or dry-coated tablets.

The tablet dosage forms can be prepared from the active ingredient inpowdered, crystalline, or granular forms, alone or in combination withone or more carriers or excipients described herein, including binders,disintegrants, controlled-release polymers, lubricants, diluents, and/orcolorants. Flavoring and sweetening agents are especially useful in theformation of chewable tablets and lozenges.

The pharmaceutical compositions provided herein can be provided as softor hard capsules, which can be made from gelatin, methylcellulose,starch, or calcium alginate. The hard gelatin capsule, also known as thedry-filled capsule (DFC), consists of two sections, one slipping overthe other, thus completely enclosing the active ingredient. The softelastic capsule (SEC) is a soft, globular shell, such as a gelatinshell, which is plasticized by the addition of glycerin, sorbitol, or asimilar polyol. The soft gelatin shells may contain a preservative toprevent the growth of microorganisms. Suitable preservatives are thoseas described herein, including methyl- and propyl-parabens, and sorbicacid. The liquid, semisolid, and solid dosage forms provided herein maybe encapsulated in a capsule. Suitable liquid and semisolid dosage formsinclude solutions and suspensions in propylene carbonate, vegetableoils, or triglycerides. Capsules containing such solutions can beprepared as described in U.S. Pat. Nos. 4,328,245; 4,409,239; and4,410,545. The capsules may also be coated as known by those of skill inthe art in order to modify or sustain dissolution of the activeingredient.

The pharmaceutical compositions provided herein can be provided inliquid and semisolid dosage forms, including emulsions, solutions,suspensions, elixirs, and syrups. An emulsion is a two-phase system, inwhich one liquid is dispersed in the form of small globules throughoutanother liquid, which can be oil-in-water or water-in-oil. Emulsions mayinclude a pharmaceutically acceptable nonaqueous liquid or solvent,emulsifying agent, and preservative. Suspensions may include apharmaceutically acceptable suspending agent and preservative. Aqueousalcoholic solutions may include a pharmaceutically acceptable acetal,such as a di(lower alkyl) acetal of a lower alkyl aldehyde, e.g.,acetaldehyde diethyl acetal; and a water-miscible solvent having one ormore hydroxyl groups, such as propylene glycol and ethanol. Elixirs areclear, sweetened, and hydroalcoholic solutions. Syrups are concentratedaqueous solutions of a sugar, for example, sucrose, and may also containa preservative. For a liquid dosage form, for example, a solution in apolyethylene glycol may be diluted with a sufficient quantity of apharmaceutically acceptable liquid carrier, e.g., water, to be measuredconveniently for administration.

Other useful liquid and semisolid dosage forms include, but are notlimited to, those containing the active ingredient(s) provided herein,and a dialkylated mono- or poly-alkylene glycol, including,1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethyleneglycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether,polyethylene glycol-750-dimethyl ether, wherein 350, 550, and 750 referto the approximate average molecular weight of the polyethylene glycol.These formulations can further comprise one or more antioxidants, suchas butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA),propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine,lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoricacid, bisulfite, sodium metabisulfite, thiodipropionic acid and itsesters, and dithiocarbamates.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The composition can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

The pharmaceutical compositions provided herein for oral administrationcan be also provided in the forms of liposomes, micelles, microspheres,or nanosystems. Micellar dosage forms can be prepared as described inU.S. Pat. No. 6,350,458.

The pharmaceutical compositions provided herein can be provided asnon-effervescent or effervescent, granules and powders, to bereconstituted into a liquid dosage form. Pharmaceutically acceptablecarriers and excipients used in the non-effervescent granules or powdersmay include diluents, sweeteners, and wetting agents. Pharmaceuticallyacceptable carriers and excipients used in the effervescent granules orpowders may include organic acids and a source of carbon dioxide.

Coloring and flavoring agents can be used in all of the above dosageforms.

The compounds disclosed herein may also be coupled with soluble polymersas targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted withpalmitoyl residues. Furthermore, the compounds disclosed herein may becoupled to a class of biodegradable polymers useful in achievingcontrolled release of a drug, for example, polylactic acid, polyepsiloncaprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,polydihydropyrans, polycyanoacrylates and cross-linked or amphipathicblock copolymers of hydrogels.

The pharmaceutical compositions provided herein can be formulated asimmediate or modified release dosage forms, including delayed-,sustained-, pulsed-, controlled-, targeted-, and programmed-releaseforms.

The pharmaceutical compositions provided herein can be co-formulatedwith other active ingredients which do not impair the desiredtherapeutic action, or with substances that supplement the desiredaction.

The pharmaceutical compositions provided herein can be administeredparenterally by injection, infusion, or implantation, for local orsystemic administration. Parenteral administration, as used herein,include intravenous, intraarterial, intraperitoneal, intrathecal,intraventricular, intraurethral, intrasternal, intracranial,intramuscular, intrasynovial, intravesical, and subcutaneousadministration.

The pharmaceutical compositions provided herein can be formulated in anydosage forms that are suitable for parenteral administration, includingsolutions, suspensions, emulsions, micelles, liposomes, microspheres,nanosystems, and solid forms suitable for solutions or suspensions inliquid prior to injection. Such dosage forms can be prepared accordingto conventional methods known to those skilled in the art ofpharmaceutical science (see, Remington: The Science and Practice ofPharmacy, supra).

The pharmaceutical compositions intended for parenteral administrationcan include one or more pharmaceutically acceptable carriers andexcipients, including, but not limited to, aqueous vehicles,water-miscible vehicles, non-aqueous vehicles, antimicrobial agents orpreservatives against the growth of microorganisms, stabilizers,solubility enhancers, isotonic agents, buffering agents, antioxidants,local anesthetics, suspending and dispersing agents, wetting oremulsifying agents, complexing agents, sequestering or chelating agents,cryoprotectants, lyoprotectants, thickening agents, pH adjusting agents,and inert gases.

Suitable aqueous vehicles include, but are not limited to, water,saline, physiological saline or phosphate buffered saline (PBS), sodiumchloride injection, Ringers injection, isotonic dextrose injection,sterile water injection, dextrose and lactated Ringers injection.Non-aqueous vehicles include, but are not limited to, fixed oils ofvegetable origin, castor oil, corn oil, cottonseed oil, olive oil,peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil,hydrogenated vegetable oils, hydrogenated soybean oil, and medium-chaintriglycerides of coconut oil, and palm seed oil. Water-miscible vehiclesinclude, but are not limited to, ethanol, 1,3-butanediol, liquidpolyethylene glycol (e.g., polyethylene glycol 300 and polyethyleneglycol 400), propylene glycol, glycerin, N-methyl-2-pyrrolidone,N,N-dimethylacetamide, and dimethyl sulfoxide.

Suitable antimicrobial agents or preservatives include, but are notlimited to, phenols, cresols, mercurials, benzyl alcohol, chlorobutanol,methyl and propyl p-hydroxybenzoates, thimerosal, benzalkonium chloride(e.g., benzethonium chloride), methyl- and propyl-parabens, and sorbicacid. Suitable isotonic agents include, but are not limited to, sodiumchloride, glycerin, and dextrose. Suitable buffering agents include, butare not limited to, phosphate and citrate. Suitable antioxidants arethose as described herein, including bisulfite and sodium metabisulfite.Suitable local anesthetics include, but are not limited to, procainehydrochloride. Suitable suspending and dispersing agents are those asdescribed herein, including sodium carboxymethylcelluose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Suitable emulsifying agentsinclude those described herein, including polyoxyethylene sorbitanmonolaurate, polyoxyethylene sorbitan monooleate 80, and triethanolamineoleate. Suitable sequestering or chelating agents include, but are notlimited to EDTA. Suitable pH adjusting agents include, but are notlimited to, sodium hydroxide, hydrochloric acid, citric acid, and lacticacid. Suitable complexing agents include, but are not limited to,cyclodextrins, including α-cyclodextrin, β-cyclodextrin,hydroxypropyl-P-cyclodextrin, sulfobutylether-β-cyclodextrin, andsulfobutylether 7-P-cyclodextrin (CAPTISOL®, CyDex, Lenexa, Kans.).

The pharmaceutical compositions provided herein can be formulated forsingle or multiple dosage administration. The single dosage formulationsare packaged in an ampoule, a vial, or a syringe. The multiple dosageparenteral formulations must contain an antimicrobial agent atbacteriostatic or fungistatic concentrations. All parenteralformulations must be sterile, as known and practiced in the art.

In one embodiment, the pharmaceutical compositions are provided asready-to-use sterile solutions. In another embodiment, thepharmaceutical compositions are provided as sterile dry solubleproducts, including lyophilized powders and hypodermic tablets, to bereconstituted with a vehicle prior to use. In yet another embodiment,the pharmaceutical compositions are provided as ready-to-use sterilesuspensions. In yet another embodiment, the pharmaceutical compositionsare provided as sterile dry insoluble products to be reconstituted witha vehicle prior to use. In still another embodiment, the pharmaceuticalcompositions are provided as ready-to-use sterile emulsions.

The pharmaceutical compositions provided herein can be formulated asimmediate or modified release dosage forms, including delayed-,sustained-, pulsed-, controlled-, targeted-, and programmed-releaseforms.

The pharmaceutical compositions can be formulated as a suspension,solid, semi-solid, or thixotropic liquid, for administration as animplanted depot. In one embodiment, the pharmaceutical compositionsprovided herein are dispersed in a solid inner matrix, which issurrounded by an outer polymeric membrane that is insoluble in bodyfluids but allows the active ingredient in the pharmaceuticalcompositions diffuse through.

Suitable inner matrixes include polymethylmethacrylate,polybutyl-methacrylate, plasticized or unplasticized polyvinylchloride,plasticized nylon, plasticized polyethylene terephthalate, naturalrubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene,ethylene-vinyl acetate copolymers, silicone rubbers,polydimethylsiloxanes, silicone carbonate copolymers, hydrophilicpolymers, such as hydrogels of esters of acrylic and methacrylic acid,collagen, cross-linked polyvinyl alcohol, and cross-linked partiallyhydrolyzed polyvinyl acetate.

Suitable outer polymeric membranes include polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinyl acetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinyl chloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer.

In another aspect, The pharmaceutical compositions disclosed herein canbe formulated in any dosage forms that are adapted for administration toa patient by inhalation, for example as a dry powder, an aerosol, asuspension, or a solution composition. In one embodiment, thepharmaceutical compositions disclosed herein can be formulated in adosage form adapted for administration to a patient by inhalation as adry powder. In a further embodiment, the pharmaceutical compositionsdisclosed herein can be formulated in a dosage form adapted foradministration to a patient by inhalation via a nebulizer. Dry powdercompositions for delivery to the lung by inhalation typically comprisethe compounds disclosed herein as a finely divided powder together withone or more pharmaceutically-acceptable excipients as finely dividedpowders. Pharmaceutically-acceptable excipients particularly suited foruse in dry powders are known to those skilled in the art and includelactose, starch, mannitol, and mono-, di-, and polysaccharides. Thefinely divided powder may be prepared by, for example, micronisation andmilling. Generally, the size-reduced (eg micronised) compound can bedefined by a D₅₀ value of about 1 to about 10 microns (for example asmeasured using laser diffraction).

Aerosols may be formed by suspending or dissolving the compounddisclosed herein in a liquified propellant. Suitable propellants includehalocarbons, hydrocarbons, and other liquified gases. Representativepropellants include: trichlorofluoromethane (propellant 11),dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane(propellant 114), tetrafluoroethane (HFA-134a), 1,1-difluoro ethane(HFA-152a), difluoromethane (HFA-32), pentafluoroethane (HFA-12),heptafluoropropane (HFA-227a), perfluoropropane, perfluorobutane,perfluoropentane, butane, isobutane, and pentane. Aerosols comprisingthe compound disclosed herein will typically be administered to apatient via a metered dose inhaler (MDI). Such devices are known tothose skilled in the art.

The aerosol may contain additional pharmaceutically-acceptableexcipients typically used with MDIs such as surfactants, lubricants,cosolvents and other excipients to improve the physical stability of theformulation, to improve valve performance, to improve solubility, or toimprove taste.

Pharmaceutical compositions adapted for transdermal administration maybe presented as discrete patches intended to remain in intimate contactwith the epidermis of the patient for a prolonged period of time. Forexample, the active ingredient may be delivered from the patch byiontophoresis as generally described in Pharmaceutical Research, 3(6),318 (1986).

Pharmaceutical compositions adapted for topical administration may beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols or oils. Ointments, creams andgels, may, for example, be formulated with an aqueous or oily base withthe addition of suitable thickening and/or gelling agent and/orsolvents. Such bases may thus, for example, include water and/or oilsuch as liquid paraffin or a vegetable oil such as arachis oil or castoroil, or a solvent such as polyethylene glycol. Thickening agents andgelling agents which may be used according to the nature of the baseinclude soft paraffin, aluminium stearate, cetostearyl alcohol,polyethylene glycols, woolfat, beeswax, carboxypolymethylene andcellulose derivatives, and/or glyceryl monostearate and/or non-ionicemulsifying agents.

Lotions may be formulated with an aqueous or oily base and will ingeneral also contain one or more emulsifying agents, stabilising agents,dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of anysuitable powder base, for example, talc, lactose or starch. Drops may beformulated with an aqueous or nonaqueous base also comprising one ormore dispersing agents, solubilizing agents, suspending agents orpreservatives.

Topical preparations may be administered by one or more applications perday to the affected area; over skin areas occlusive dressings mayadvantageously be used. Continuous or prolonged delivery may be achievedby an adhesive reservoir system.

For treatments of the eye or other external tissues, for example mouthand skin, the compositions may be applied as a topical ointment orcream. When formulated in an ointment, the compound disclosed herein maybe employed with either a paraffinic or a water-miscible ointment base.Alternatively, the compound disclosed herein may be formulated in acream with an oil-in-water cream base or a water-in-oil base.

Use of the Compounds and Compositions of the Invention

The present invention provides a method of using a compound disclosedherein, or a pharmaceutical composition comprising the compounddisclosed herein for the treatment, prevention, or amelioration of adisease or disorder that is mediated or otherwise affected via one ormore protein kinases activiy, such as JAK kinase (including JAK1, JAK2,JAK3 or TYK2 kinase), FLT3 kinase, and Aurora kinase (includingAurora-A, Aurora-B and Aurora C) activiy or one or more symptoms ofdiseases or disorders that are mediated or otherwise affected via one ormore protein kinases activiy, such as JAK kinase (including JAK1, JAK2,JAK3 or TYK2 kinase), FLT3 kinase and Aurora kinase (including Aurora-A,Aurora-B and Aurora C kinase) activity.

FLT3 kinase can be wild type and/or mutant form of FLT3 kinase.

JAK kinase can be wild type and/or mutant form of JAK1, JAK2, JAK3 orTYK2 kinase.

In one embodiment, provided herein is a method of using a compounddisclosed herein or a pharmaceutical composition comprising a compounddisclosed herein for the treatment, prevention, or amelioration of adisease or disorder that is mediated or otherwise affected viainappropriate JAK1 kinase activity or one or more symptoms of diseasesor disorders that are mediated or otherwise affected via inappropriateJAK1 kinase activity. In another embodiment, a disease, a disorder orone or more symptoms of diseases or disorders is related to theinappropriate activity of JAK2 kinase. In yet another embodiment, adisease, a disorder or one or more symptoms of diseases or disorders isrelated to the inappropriate activity of JAK3 kinase.

In one embodiment, provided herein is a method of using a compounddisclosed herein or a pharmaceutical composition comprising a compounddisclosed herein for the treatment, prevention, or amelioration of adisease or disorder that is mediated or otherwise affected viainappropriate FLT3 kinase activity or one or more symptoms of diseasesor disorders that are mediated or otherwise affected via inappropriateFLT3 kinase activity.

In one embodiment, provided herein is a method of using a compounddisclosed herein or a pharmaceutical composition comprising a compounddisclosed herein for the treatment, prevention, or amelioration of adisease or disorder that is mediated or otherwise affected viainappropriate Aurora-A kinase activity or one or more symptoms ofdiseases or disorders that are mediated or otherwise affected viainappropriate Aurora-A kinase activity. In another embodiment, adisease, a disorder or one or more symptoms of diseases or disorders isrelated to the inappropriate activity of Aurora-B kinase. In yet anotherembodiment, a disease, a disorder or one or more symptoms of diseases ordisorders is related to the inappropriate activity of Aurora C kinase.

“Inappropriate JAK kinase activity” refers to any JAK kinase activitythat deviates from the normal JAK kinase activity expected in aparticular patient. Inappropriate JAK kinase may take the form of, forinstance, an abnormal increase in activity, or an aberration in thetiming and or control of JAK kinase activity. Such inappropriateactivity may result then, for example, from overexpression or mutationof the protein kinase leading to inappropriate or uncontrolledactivation. Accordingly, in another aspect the invention is directed tomethods of treating such diseases and disorders.

Consistent with the description above, such diseases or disordersinclude without limitation: myeloproliferative disorders such aspolycythemia vera (PCV), essential thrombocythemia and idiopathicmyelofibrosis (IMF); leukemia such as myeloid leukemia including chronicmyeloid leukemia (CML), imatinib-resistant forms of CML, acute myeloidleukemia (AML), and a subtype of AML, acute megakaryoblastic leukemia(AMKL); lymphoproliferative diseases such as acute lymphocytic leukemia(ALL) and myeloma; cancer including head and neck cancer, prostatecancer, breast cancer, ovarian cancer, melanoma, lung cancer, braintumor, pancreatic cancer and renal carcinoma; and allergic orinflammatory diseases or disorders related to immune dysfunction,immunodeficiency, immunomodulation, autoimmune diseases, transplantationrejection, graft-versus-host disease, wound healing, kidney disease,multiple sclerosis, thyroiditis, type 1 diabetes, sarcoidosis,psoriasis, allergic rhinitis, inflammatory bowel disease includingCrohn's disease and ulcerative colitis (UC), systemic lupuserythematosis (SLE), arthritis, osteoarthritis, rheumatoid arthritis,osteoporosis, asthma and chronic obstructive pulmonary disease (COPD)and dry eye syndrome (or keratoconjunctivitis sicca (KCS)).

In one aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for preventing and/or treatingproliferative disease, autoimmune disease, allergic disease,inflammatory disease or transplantation rejection in mammals includinghumans.

In yet another aspect, provided herein is a method of treating a mammalhaving, or at risk of having a disease or disclosed herein, said methodcomprising administering an effective condition-treating orcondition-preventing amount of one or more of the pharmaceuticalcompositions or the compounds disclosed herein. In a particular aspect,provided here is a method of treating a mammal having, or at risk ofhaving proliferative disease, autoimmune disease, allergic disease,inflammatory disease or transplantation rejection.

In additional method of treatment aspects, provided herein is a methodof treatment and/or prophylaxis of a mammal susceptible to or afflictedwith a proliferative disease, said methods comprising administering aneffective condition-treating or condition-preventing amount of one ormore of the pharmaceutical compositions or compounds disclosed herein.In a specific embodiment, the proliferative disease is selected fromcancer (e.g. solid tumors such as uterine leiomyosarcoma or prostatecancer), polycythemia vera, essential thrombocytosis, myelofibrosis,leukemia (e.g. AML, CML, ALL or CLL), and multiple myeloma.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment, and/orprophylaxis of a proliferative disease. In a specific embodiment, theproliferative disease is selected from cancer (e.g. solid tumors such asuterine leiomyosarcoma or prostate cancer), polycythemia vera, essentialthrombocytosis, myelofibrosis, leukemia (e.g. AML, CML, ALL or CLL), andmultiple myeloma.

In yet another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein for use in the manufactureof a medicament for the treatment, and/or prophylaxis of a proliferativedisease. In a specific embodiment, the proliferative disease is selectedfrom cancer (e.g. solid tumors such as uterine leiomyosarcoma orprostate cancer), polycythemia vera, essential thrombocytosis,myelofibrosis, leukemia (e.g. AML, CML, ALL or CLL), and multiplemyeloma.

In another aspect, provided herein is a method of treatment and/orprophylaxis of a mammal susceptible to or afflicted with an autoimmunedisease. The methods comprise administering an effectivecondition-treating or condition-preventing amount of one or more of thepharmaceutical compositions or compounds disclosed herein. In a specificembodiment, the autoimmune disease is selected from COPD, asthma,systemic and cutaneous lupus erythematosis, lupus nephritis,dermatomyositis, Sjogren's syndrome, psoriasis and type I diabetesmellitus.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment, and/orprophylaxis of an autoimmune disease. In a specific embodiment, theautoimmune disease is selected from COPD, asthma, systemic and cutaneouslupus erythematosis, lupus nephritis, dermatomyositis, Sjogren'ssyndrome, psoriasis and type I diabetes mellitus.

In yet another aspect, provided here is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for the treatment, and/or prophylaxis of an autoimmunedisease. In a specific embodiment, the autoimmune disease is selectedfrom COPD, asthma, systemic and cutaneous lupus erythematosis, lupusnephritis, dermatomyositis, Sjogren's syndrome, psoriasis and type Idiabetes mellitus.

In a method of treatment aspects, provided herein are methods oftreatment and/or prophylaxis of a mammal susceptible to or afflictedwith an allergic disease. The methods comprising administering aneffective condition-treating or condition-preventing amount of one ormore of the pharmaceutical compositions or the compounds disclosedherein. In a specific embodiment, the allergic disease is selected fromallergic airway disease, sinusitis, eczema and hives, food allergies andallergies to insect venom.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment, and/orprophylaxis of an allergic disease. In a specific embodiment, theallergic disease is selected from allergic airway disease, sinusitis,eczema and hives, food allergies and allergies to insect venom.

In yet another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for the treatment, or prophylaxis of an allergic disease. Ina specific embodiment, the allergic disease is selected from allergicairway disease, sinusitis, eczema and hives, food allergies andallergies to insect venom.

In another aspect, provided herein are methods of treatment and/orprophylaxis of a mammal susceptible to or afflicted with an inflammatorydisease. The methods comprise administering an effectivecondition-treating or condition-preventing amount of one or more of thepharmaceutical compositions or the compounds disclosed herein. In aspecific embodiment, the inflammatory disease is selected frominflammatory bowel syndrome, Crohn's disease, rheumatoid arthritis,juvenile arthritis and psoriatic arthritis.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment, and/orprophylaxis of an inflammatory disease. In a specific embodiment, theinflammatory disease is selected from inflammatory bowel syndrome,Crohn's disease, rheumatoid arthritis, juvenile arthritis and psoriaticarthritis.

In yet another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for the treatment, and/or prophylaxis of an inflammatorydisease. In a specific embodiment, the inflammatory disease is selectedfrom inflammatory bowel syndrome, Crohn's disease, rheumatoid arthritis,juvenile arthritis and psoriatic arthritis.

In another aspect, provided herein are methods of treatment and/orprophylaxis of a mammal susceptible to or afflicted with transplantationrejection. The methods comprising administering an effectivecondition-treating or condition-preventing amount of one or more of thepharmaceutical compositions or the compound of the invention hereindescribed. In a specific embodiment, the transplantation rejection isorgan transplant rejection, tissue transplant rejection and celltransplant rejection.

In another aspect, provided herein is the compound or the pharmaceuticalcomposition disclosed herein for use in the treatment, and/orprophylaxis of transplantation rejection. In a specific embodiment, thetransplantation rejection is organ transplant rejection, tissuetransplant rejection and cell transplant rejection.

In yet another aspect, provided herein is the use of the compound or thepharmaceutical composition disclosed herein for use in the manufactureof a medicament for the treatment and/or prophylaxis of transplantationrejection. In a specific embodiment, the transplantation rejection isorgan transplant rejection, tissue transplant rejection and celltransplant rejection.

The present invention provides the compound or the pharmaceuticalcomposition disclosed herein for use as a pharmaceutical especially inthe treatment and/or prophylaxis of the aforementioned diseases ordisorders. Also provided herein is the use of the compound or thepharmaceutical composition disclosed herein in the manufacture of amedicament for the treatment and/or prophylaxis of one of theaforementioned diseases or disorders.

A particular regimen of the present method comprises the administrationto a subject suffering from a disease involving inflammation, of aneffective amount of a compound disclosed herein for a period of timesufficient to reduce the level of inflammation in the subject, andpreferably terminate the processes responsible for said inflammation. Aspecial embodiment of the method comprises administering of an effectiveamount of a compound disclosed herein to a subject patient sufferingfrom or susceptible to the development of rheumatoid arthritis, for aperiod of time sufficient to reduce or prevent, respectively,inflammation in the joints of said patient, and preferably terminate,the processes responsible for said inflammation.

A further particular regimen of the present method comprises theadministration to a subject suffering from a disease involvingproliferative disease, of an effective amount of a compound disclosedherein for a period of time sufficient to reduce the level ofproliferative disease in the subject, and preferably terminate theprocesses responsible for said proliferative disease. A particularembodiment of the method comprises administering of an effective amountof a compound disclosed herein to a subject patient suffering from orsusceptible to the development of cancer, for a period of timesufficient to reduce or prevent, respectively, solid tumor of saidpatient, and preferably terminate, the processes responsible for saidsolid.

Combination Therapy

A compound disclosed herein can be administered as the sole active agentor it can be administered in combination with other therapeutic agents,including other compounds that demonstrate the same or a similartherapeutic activity and that are determined to be safe and efficaciousfor such combined administration.

In one aspect, provided herein is a method of treating, preventing, orameliorating a disease or disorder comprising administering a safe andeffective amount of a combination comprising the compound disclosedherein together with one or more therapeutically active agents. In oneembodiment, the combinations comprising one or two other therapeuticagents.

Example of other therapeutic agents may include without limitationanti-cancer agents, including chemotherapeutic agents andantiproliferative agents; anti-inflammatory agents and immunomodulatoryagents or immunosuppressive agents.

In another aspect, provided herein is a product comprising a compounddisclosed herein and at least one other therapeutic agent as a combinedpreparation for simultaneous, separate or sequential use in therapy. Inone embodiment, the therapy is the treatment of a disease or disordermediated by the activity of one or more protein kinases activiy, such asJAK kinase, FLT3 kinase and Aurora kinase. Products provided as acombined preparation include a composition comprising the compounddisclosed herein and the other therapeutic agent(s) together in the samepharmaceutical composition, or the compound disclosed herein and theother therapeutic agent(s) in separate form, e.g. in the form of a kit.

In another aspect, provided herein is a pharmaceutical compositioncomprising a compound disclosed herein and another therapeutic agent(s).In one embodiment, the pharmaceutical composition may comprise apharmaceutically acceptable excipient, carrier, adjuvant or veichle asdescribed above.

In another aspect, the invention provides a kit comprising two or moreseparate pharmaceutical compositions, at least one of which contains acompound disclosed herein. In one embodiment, the kit comprises meansfor separately retaining said compositions, such as a container, dividedbottle, or divided foil packet. An example of such a kit is a blisterpack, as typically used for the packaging of tablets, capsules and thelike.

The invention also provides the use of a compound disclosed herein fortreating a disease or condition mediated by the activity of one or moreprotein kinases, such as JAK kinase, FLT3 kinase and Aurora kinase,wherein the patient has previously (e.g. within 24 hours) been treatedwith another therapeutic agent. The invention also provides the use ofanother therapeutic agent for treating a disease or condition mediatedby the activity of one or more protein kinases, such as JAK kinase, FLT3kinase and Aurora kinase, wherein the patient has previously (e.g.within 24 hours) been treated with a compound disclosed herein.

The compounds disclosed herein may be administered as the sole activeingredient or in conjunction with, e.g. as an adjuvant to, other drugse.g. immunosuppressive or immunomodulating agents or otheranti-inflammatory agents, e.g. for the treatment or prevention of alio-or xenograft acute or chronic rejection or inflammatory or autoimmunedisorders, or a chemotherapeutic agent, e.g a malignant cellanti-proliferative agent. For example, the compounds disclosed hereinmay be used in combination with a calcineurin inhibitor, e.g.cyclosporin A or FK 506; a mTOR inhibitor, e.g. rapamycin,40-O-(2-hydroxyethyl)-rapamycin, CCI779, ABT578, AP23573, TAFA-93,biolimus-7 or biolimus-9; an ascomycin having immuno-suppressiveproperties, e.g. ABT-281, ASM981, etc.; corticosteroids;cyclophosphamide; azathioprene; methotrexate; leflunomide; mizoribine;mycophenolic acid or salt; mycophenolate mofetil; 15-deoxyspergualine oran immunosuppressive homologue, analogue or derivative thereof; a PKCinhibitor, e.g. as disclosed in WO 02/38561 or WO 03/82859, e.g. thecompound of Example 56 or 70; immunosuppressive monoclonal antibodies,e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3,CD4, CD7, CD8, CD25, CD28, CD40, CD45, CD52, CD58, CD80, CD86 or theirligands; other immunomodulatory compounds, e.g. a recombinant bindingmolecule having at least a portion of the extracellular domain of CTLA4or a mutant thereof, e.g. an at least extracellular portion of CTLA4 ora mutant thereof joined to a non-CTLA4 protein sequence, e.g. CTLA41g(for ex. designated ATCC 68629) or a mutant thereof, e.g. LEA29Y;adhesion molecule inhibitors, e.g. LFA-1 antagonists, ICAM-1 or -3antagonists, VCAM-4 antagonists or VLA-4 antagonists; or antihistamines;or antitussives, or a bronchodilatory agent; or an angiotensin receptorblockers; or an anti-infectious agent.

Where the compounds disclosed herein are administered in conjunctionwith other immunosuppressive/immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious therapy, dosages of theco-administered immunosuppressant, immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious compound will of course varydepending on the type of co-drug employed, e.g. whether it is a steroidor a calcineurin inhibitor, on the specific drug employed, on thecondition being treated and so forth.

In one aspect, provided herein is a combination comprising a compounddisclosed herein together with a β₂-adrenoreceptor agonist. Examples ofβ₂-adrenoreceptor agonists include salmeterol, salbutamol, formoterol,salmefamol, fenoterol, carmoterol, etanterol, naminterol, clenbuterol,pirbuterol, flerbuterol, reproterol, bambuterol, indacaterol,terbutaline and salts thereof, for example the xinafoate(1-hydroxy-2-naphthalenecarboxylate) salt of salmeterol, the sulphatesalt or free base of salbutamol or the fumarate salt of formoterol. Inone embodiment, long-acting β₂-adrenoreceptor agonists, for example,compounds which provide effective bronchodilation for about 12 h orlonger, are preferred.

The β₂-adrenoreceptor agonist may be in the form of a salt formed with apharmaceutically acceptable acid selected from sulphuric, hydrochloric,fumaric, hydroxynaphthoic (for example 1- or 3-hydroxy-2-naphthoic),cinnamic, substituted cinnamic, triphenylacetic, sulphamic, sulphanilic,naphthaleneacrylic, benzoic, 4-methoxybenzoic, 2- or 4-hydroxybenzoic,4-chlorobenzoic and 4-phenylbenzoic acid.

In another aspect, provided herein is a combination comprising acompound disclosed herein together with corticosteroids. Suitablecorticosteroids refer to those oral and inhaled corticosteroids andtheir pro-drugs which have anti-inflammatory activity. Examples includemethyl prednisolone, prednisolone, dexamethasone, fluticasonepropionate,6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-[(4-methyl-1,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester,6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester (fluticasone furoate),6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-propionyloxy-androsta-1,4-diene-17β-carbothioicacid S-(2-oxo-tetrahydro-furan-3S-yl) ester,6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-(2,2,3,3-tetramethycyclopropylcarbonyl)oxy-androsta-1,4-diene-17β-carbothioicacid S-cyanomethyl ester and6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-(1-ethycyclopropylcarbonyl)oxy-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester, beclomethasone esters (for example the17-propionate ester or the 17,21-dipropionate ester), budesonide,flunisolide, mometasone esters (for example mometasone furoate),triamcinolone acetonide, rofleponide, ciclesonide(16α,17-[[(cis)-cyclohexylmethylene]bis(oxy)]-11β,21-dihydroxy-pregna-1,4-diene-3,20-dione),butixocort propionate, RPR-106541, and ST-126. Preferred corticosteroidsinclude fluticasone propionate,6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-[(4-methyl-1,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester,6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester,6α,9α-difluoro-11β-hydroxy-16α-methyl-3-oxo-17α-(2,2,3,3-tetramethycyclopropylcarbonyl)oxy-androsta-1,4-diene-17β-carbothioicacid S-cyanomethyl ester and6α,9α-difluoro-11β-hydroxy-16α-methyl-17α-(1-methylcyclopropylcarbonyl)oxy-3-oxo-androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester. In one embodiment the corticosteroid is6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester.

In another aspect, provided herein is a combination comprising acompound disclosed herein together with non-steroidal GR agonist.Non-steroidal compounds having glucocorticoid agonism that may possessselectivity for transrepression over transactivation and that may beuseful in combination therapy include those covered in the followingpatents: WO 03/082827, WO 98/54159, WO 04/005229, WO 04/009017, WO04/018429, WO 03/104195, WO 03/082787, WO 03/082280, WO 03/059899, WO03/101932, WO 02/02565, WO 01/16128, WO 00/66590, WO 03/086294, WO04/026248, WO 03/061651 and WO 03/08277. Further non-steroidal compoundsare covered in: WO 2006/000401, WO 2006/000398 and WO 2006/015870.

In another aspect, provided herein is a combination comprising acompound disclosed herein together with non-steroidal anti-inflammatorydrugs (NSAID's). Examples of NSAID's include sodium cromoglycate,nedocromil sodium, phosphodiesterase (PDE) inhibitors (for example,theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors),leukotriene antagonists, inhibitors of leukotriene synthesis (forexample montelukast), iNOS inhibitors, tryptase and elastase inhibitors,beta-2 integrin antagonists and adenosine receptor agonists orantagonists (e.g. adenosine 2a agonists), cytokine antagonists (forexample chemokine antagonists, such as a CCR3 antagonist) or inhibitorsof cytokine synthesis, or 5-lipoxygenase inhibitors. An iNOS (induciblenitric oxide synthase inhibitor) is preferably for oral administration.Examples of iNOS inhibitors include those disclosed in WO 93/13055, WO98/30537, WO 02/50021, WO 95/34534 and WO 99/62875. Examples of CCR3inhibitors include those disclosed in WO 02/26722.

In one embodiment, the invention provides the use of the compoundsdisclosed herein in combination with a phosphodiesterase 4 (PDE4)inhibitor, especially in the case of a formulation adapted forinhalation. The PDE4-specific inhibitor useful in this aspect of theinvention may be any compound that is known to inhibit the PDE4 enzymeor which is discovered to act as a PDE4 inhibitor, and which are onlyPDE4 inhibitors, not compounds which inhibit other members of the PDEfamily, such as PDE3 and PDE5, as well as PDE4. Compounds includecis-4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexan-1-carboxylicacid,2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-oneandcis-[4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-ol].Also,cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexane-1-carboxylicacid (also known as cilomilast) and its salts, esters, pro-drugs orphysical forms, which is described in U.S. Pat. No. 5,552,438 issued 3Sep. 1996; this patent and the compounds it discloses are incorporatedherein in full by reference.

In another aspect, provided herein is a combination comprising acompound disclosed herein together with an anticholinergic agent.Examples of anticholinergic agents are those compounds that act asantagonists at the muscarinic receptors, in particular those compoundswhich are antagonists of the M₁ or M₃ receptors, dual antagonists of theM₁/M₃ or M₂/M₃, receptors or pan-antagonists of the M₁/M₂/M₃ receptors.Exemplary compounds for administration via inhalation includeipratropium (for example, as the bromide, CAS 22254-24-6, sold under thename ATROVENT®), oxitropium (for example, as the bromide, CAS30286-75-0) and tiotropium (for example, as the bromide, CAS136310-93-5, sold under the name SPIRIVA®). Also of interest arerevatropate (for example, as the hydrobromide, CAS 262586-79-8) andLAS-34273 which is disclosed in WO 01/04118. Exemplary compounds fororal administration include pirenzepine (CAS 28797-61-7), darifenacin(CAS 133099-04-4, or CAS 133099-07-7 for the hydrobromide sold under thename ENABLEX®), oxybutynin (CAS 5633-20-5, sold under the nameDITROPAN®), terodiline (CAS 15793-40-5), tolterodine (CAS 124937-51-5,or CAS 124937-52-6 for the tartrate, sold under the name DETROL®),otilonium (for example, as the bromide, CAS 26095-59-0, sold under thename SPASMOMEN®), trospium chloride (CAS 10405-02-4) and solifenacin(CAS 242478-37-1, or CAS 242478-38-2 for the succinate also known asYM-905 and sold under the name VESICARE®).

In another aspect, provided herein is a combination comprising acompound disclosed herein together with an H1 antagonist. Examples of H1antagonists include, without limitation, amelexanox, astemizole,azatadine, azelastine, acrivastine, brompheniramine, cetirizine,levocetirizine, efletirizine, chlorpheniramine, clemastine, cyclizine,carebastine, cyproheptadine, carbinoxamine, descarboethoxyloratadine,doxylamine, dimethindene, ebastine, epinastine, efletirizine,fexofenadine, hydroxyzine, ketotifen, loratadine, levocabastine,mizolastine, mequitazine, mianserin, noberastine, meclizine,norastemizole, olopatadine, picumast, pyrilamine, promethazine,terfenadine, tripelennamine, temelastine, trimeprazine and triprolidine,particularly cetirizine, levocetirizine, efletirizine and fexofenadine.In a further embodiment the invention provides a combination comprisinga compound disclosed herein together with an H3 antagonist (and/orinverse agonist). Examples of H3 antagonists include, for example, thosecompounds disclosed in WO 2004/035556 and in WO 2006/045416. Otherhistamine receptor antagonists which may be used in combination with thecompounds disclosed herein include antagonists (and/or inverse agonists)of the H4 receptor, for example, the compounds disclosed in Jablonowskiet al., J. Med. Chem. 46:3957-3960 (2003).

In still another aspect, provided herein is a combination comprising acompound disclosed herein together with a PDE4 inhibitor and aβ₂-adrenoreceptor agonist.

In yet another aspect, provided herein is a combination comprising acompound disclosed herein together with an anticholinergic and a PDE-4inhibitor.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical composition and thus pharmaceuticalcompositions comprising a combination as defined above together with apharmaceutically acceptable excipient or carrier represent a furtheraspect of the invention.

The individual compounds of such combinations may be administered eithersequentially or simultaneously in separate or combined pharmaceuticalformulations. In one embodiment, the individual compounds will beadministered simultaneously in a combined pharmaceutical formulation.Appropriate doses of known therapeutic agents will readily beappreciated by those skilled in the art.

The invention thus provides, in a further aspect, a pharmaceuticalcomposition comprising a combination of a compound disclosed hereintogether with another therapeutically active agent.

In one embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with a PDE4inhibitor.

In another embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with aβ₂-adrenoreceptor agonist.

In another embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with acorticosteroid.

In another embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with a non-steroidalGR agonist.

In another embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with ananticholinergic agent.

In still another embodiment, the pharmaceutical composition comprises acombination of a compound disclosed herein together with anantihistamine.

In the field of medical oncology it is normal practice to use acombination of different forms of treatment to treat each patient withcancer. In medical oncology the other component(s) of such conjointtreatment in addition to compositions disclosed herein may be, forexample, surgery, radiotherapy, chemotherapy, signal transductioninhibitors or modulators (e.g. kinase inhibitors or modulators) and/ormonoclonoal antibodies.

A compound disclosed herein may also be used to advantage in combinationwith each other or in combination with other therapeutic agents,especially other antiproliferative agents. Such antiproliferative agentsinclude, but are not limited to, aromatase inhibitors; antiestrogens;topoisomerase I inhibitors; topoisomerase II inhibitors; microtubuleactive agents; alkylating agents; histone deacetylase inhibitors;compounds that induce cell differentiation processes; cyclooxygenaseinhibitors; MMP inhibitors; mTOR inhibitors; antineoplasticantimetabolites; platin compounds; compounds targeting/decreasing aprotein or lipid kinase activity and further anti-angiogenic compounds;compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase; gonadorelin agonists; anti-androgens; methionineaminopeptidase inhibitors; bisphosphonates; biological responsemodifiers; antiproliferative antibodies; heparanase inhibitors;inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasomeinhibitors; agents used in the treatment of hematologic malignancies;compounds which target, decrease or inhibit the activity of Flt-3; Hsp90inhibitors; temozolomide (TEMODAL®); and leucovorin.

The term “aromatase inhibitor”, as used herein, relates to a compoundwhich inhibits the estrogen production, i.e., the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to, steroids,especially atamestane, exemestane and formestane; and, in particular,nonsteroids, especially aminoglutethimide, roglethimide,pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole,fadrozole, anastrozole and letrozole. Exemestane can be administered,e.g., in the form as it is marketed, e.g., under the trademarkAROMASIN®. Formestane can be administered, e.g., in the form as it ismarketed, e.g., under the trademark LENTARON®. Fadrozole can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark AFEMA®. Anastrozole can be administered, e.g., in the form asit is marketed, e.g., under the trademark ARIMIDEX®. Letrozole can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark FEMARA® or FEMAR®. Aminoglutethimide can be administered,e.g., in the form as it is marketed, e.g., under the trademarkORIMETEN®. A combination of the invention comprising a chemotherapeuticagent which is an aromatase inhibitor is particularly useful for thetreatment of hormone receptor positive tumors, e.g., breast tumors.

The term “anti-estrogen”, as used herein, relates to a compound whichantagonizes the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to, tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen can be administered, e.g., inthe form as it is marketed, e.g., under the trademark NOLVADEX®.Raloxifene hydrochloride can be administered, e.g., in the form as it ismarketed, e.g., under the trademark EVISTA®. Fulvestrant can beformulated as disclosed in U.S. Pat. No. 4,659,516 or it can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark FASLODEX®. A combination of the invention comprising achemotherapeutic agent which is an antiestrogen is particularly usefulfor the treatment of estrogen receptor positive tumors, e.g., breasttumors.

The term “anti-androgen”, as used herein, relates to any substance whichis capable of inhibiting the biological effects of androgenic hormonesand includes, but is not limited to, bicalutamide (CASODEX®), which canbe formulated, e.g., as disclosed in U.S. Pat. No. 4,636,505.

The term “gonadorelin agonist”, as used herein, includes, but is notlimited to, abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., inthe form as it is marketed, e.g., under the trademark ZOLADEX®. Abarelixcan be formulated, e.g., as disclosed in U.S. Pat. No. 5,843,901. Theterm “topoisomerase I inhibitor”, as used herein, includes, but is notlimited to, topotecan, gimatecan, irinotecan, camptothecian and itsanalogues, 9-nitrocamptothecin and the macromolecular camptothecinconjugate PNU-166148 (compound A1 in WO 99/17804). Irinotecan can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark CAMPTOSAR®. Topotecan can be administered, e.g., in the formas it is marketed, e.g., under the trademark HYCAMTIN®.

The term “topoisomerase II inhibitor”, as used herein, includes, but isnot limited to, the anthracyclines, such as doxorubicin, includingliposomal formulation, e.g., CAELYX®; daunorubicin; epirubicin;idarubicin; nemorubicin; the anthraquinones mitoxantrone andlosoxantrone; and the podophillotoxines etoposide and teniposide.Etoposide can be administered, e.g., in the form as it is marketed,e.g., under the trademark ETOPOPHOS®. Teniposide can be administered,e.g., in the form as it is marketed, e.g., under the trademark VM26-BRISTOL®. Doxorubicin can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ADRIBLASTIN® or ADRIAMYCIN®.

Epirubicin can be administered, e.g., in the form as it is marketed,e.g., under the trademark FARMORUBICIN®. Idarubicin can be administered,e.g., in the form as it is marketed, e.g., under the trademark ZAVEDOS®.Mitoxantrone can be administered, e.g., in the form as it is marketed,e.g., under the trademark NOVANTRON®.

The term “microtubule active agent” relates to microtubule stabilizing,microtubule destabilizing agents and microtublin polymerizationinhibitors including, but not limited to, taxanes, e.g., paclitaxel anddocetaxel; vinca alkaloids, e.g., vinblastine, especially vinblastinesulfate; vincristine, especially vincristine sulfate and vinorelbine;discodermolides; cochicine; and epothilones and derivatives thereof,e.g., epothilone B or D or derivatives thereof. Paclitaxel may beadministered, e.g., in the form as it is marketed, e.g., TAXOL®.Docetaxel can be administered, e.g., in the form as it is marketed,e.g., under the trademark TAXOTERE®. Vinblastine sulfate can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark VINBLASTIN R.P®. Vincristine sulfate can be administered,e.g., in the form as it is marketed, e.g., under the trademarkFARMISTIN®. Discodermolide can be obtained, e.g., as disclosed in U.S.Pat. No. 5,010,099. Also included are epothilone derivatives which aredisclosed in WO 98/10121, U.S. Pat. No. 6,194,181, WO 98/25929, WO98/08849, WO 99/43653, WO 98/22461 and WO 00/31247. Especially preferredare epothilone A and/or B.

The term “alkylating agent”, as used herein, includes, but is notlimited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNUor Gliadel). Cyclophosphamide can be administered, e.g., in the form asit is marketed, e.g., under the trademark CYCLOSTIN®. Ifosfamide can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark HOLOXAN®.

The term “histone deacetylase inhibitors” or “MAC inhibitors” relates tocompounds which inhibit the histone deacetylase and which possessantiproliferative activity. This includes compounds disclosed in WO02/22577, especiallyN-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-properiamide,N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamideand pharmaceutically acceptable salts thereof. It further especiallyincludes suberoylanilide hydroxamic acid (SAHA).

The term “antineoplastic antimetabolite” includes, but is not limitedto, 5-fluorouracil or 5-FU; capecitabine; gemcitabine; DNA demethylatingagents, such as 5-azacytidine and decitabine; methotrexate andedatrexate; and folic acid antagonists, such as pemetrexed. Capecitabinecan be administered, e.g., in the form as it is marketed, e.g., underthe trademark XELODA®. Gemcitabine can be administered, e.g., in theform as it is marketed, e.g., under the trademark GEMZAR®. Also includedis the monoclonal antibody trastuzumab which can be administered, e.g.,in the form as it is marketed, e.g., under the trademark HERCEPTIN®.

The term “platin compound”, as used herein, includes, but is not limitedto, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatincan be administered, e.g., in the form as it is marketed, e,g., underthe trademark CARBOPLAT®, Oxaliplatin can be administered, e,g., in theform as it is marketed, e.g., under the trademark ELOXATIN®.

The term “compounds targeting/decreasing a protein or lipid kinaseactivity; or a protein or lipid phosphatase activity; or furtheranti-angiogenic compounds”, as used herein, includes, but is not limitedto, protein tyrosine kinase and/or serine and/or threonine kinaseinhibitors or lipid kinase inhibitors, e.g.,

a) compounds targeting, decreasing or inhibiting the activity of theplatelet-derived growth factor-receptors (PDGFR), such as compoundswhich target, decrease or inhibit the activity of PDGFR, especiallycompounds which inhibit the PDGF receptor, e,g., aN-phenyl-2-pyrimidine-amine derivative, e.g., imatinib, SU101, SU6668and GFB-111;

b) compounds targeting, decreasing or inhibiting the activity of thefibroblast growth factor-receptors (EGFR);

c) compounds targeting, decreasing or inhibiting the activity of theinsulin-like growth factor receptor I (IGF-IR), such as compounds whichtarget, decrease or inhibit the activity of IGF-IR, especially compoundswhich inhibit the receptor, such as those compounds disclosed in WO02/092599;

d) compounds targeting, decreasing or inhibiting the activity of the Trkreceptor tyrosine kinase family;

e) compounds targeting, decreasing or inhibiting the activity of the Axlreceptor tyrosine kinase family;

f) compounds targeting, decreasing or inhibiting the activity of thec-Met receptor;

g) compounds targeting, decreasing or inhibiting the activity of theKit/SCFR receptor tyrosine kinase;

h) compounds targeting, decreasing or inhibiting the activity of thec-kit receptor tyrosine kinases—(part of the PDGFR family), such ascompounds which target, decrease or inhibit the activity of the c-Kitreceptor tyrosine kinase family, especially compounds which inhibit thec-Kit receptor, e.g., imatinib;

i) compounds targeting, decreasing or inhibiting the activity of membersof the c-Abl family and their gene-fusion products, e.g., BCR-Ablkinase, such as compounds which target decrease or inhibit the activityof c-Abl family members and their gene fusion products, e.g., a.N-phenyl-2-pyrimidine-amine derivative, e.g., imatinib, PD180970, AG957,NSC 680410 or PD173955 from ParkeDavis;

j) compounds targeting, decreasing or inhibiting the activity of membersof the protein kinase C (PKC) and Raf family of serine/threoninekinases, members of the MEK, SRC, JAK, FAK, PDK and Ras/MAPK familymembers, or PI(3) kinase family, or of the PI(3)-kinase-related kinasefamily, and/or members of the cyclin-dependent kinase family (CDK) andare especially those staurosporine derivatives disclosed in U.S. Pat.No. 5,093,330, e.g., midostaurin; examples of further compounds include,e.g., UCN-01; safingol; BAY 43-9006; Bryostatin I; Perifosine;Ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521;LY333531/LY379196; isochinoline compounds, such as those disclosed in WO00/09495; FTIs; PD184352; or QAN697 (a PI3K inhibitor);

k) compounds targeting, decreasing or inhibiting the activity ofprotein-tyrosine kinase inhibitors, such as compounds which target,decrease or inhibit the activity of protein-tyrosine kinase inhibitorsinclude imatinib mesylate (GLEEVEC®) or tyrphostin. A tyrphostin ispreferably a low molecular weight (Mr<1500) compound, or apharmaceutically acceptable salt thereof, especially a compound selectedfrom the benzylidenemalonitrile class or the S-arylbenzenemalonirile orbisubstrate quinoline class of compounds, more especially any compoundselected from the group consisting of Tyrphostin A23/RG-50810, AG 99,Tyrphostin AG 213, Tyrphostin AG 1748, Tyrphostin AG 490, TyrphostinB44, Tyrphostin B44 (+) enantiomer, Tyrphostin AG 555, AG 494,Tyrphostin AG 556, AG957 and adaphostin(4-{(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester, NSC680410, adaphostin; and

l) compounds targeting, decreasing or inhibiting the activity of theepidermal growth factor family of receptor tyrosine kinases (EGFR,ErbB2, ErbB3, ErbB4 as homo- or hetero-dimers), such as compounds whichtarget, decrease or inhibit the activity of the epidermal growth factorreceptor family are especially compounds, proteins or antibodies whichinhibit members of the EGF receptor tyrosine kinase family, e.g., EGFreceptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF related ligands,and are in particular those compounds, proteins or monoclonal antibodiesgenerically and specifically disclosed in WO 97/02266, e.g., thecompound of Example 39, or in EP 0564409; WO 99/03854; EP 0520722; EP0566226; EP 0787722; EP 0837063; U.S. Pat. No. 5,747,498; WO 98/10767;WO 97/30034; WO 97/49688; WO 97/38983 and, especially, WO 96/30347,e.g., compound known as CP 358774; WO 96/33980, e.g., compound ZD 1839;and WO 95/03283, e.g., compound ZM105180, e.g., trastuzumab (HERCEPTIN),cetuximab, Iressa, Tarceva, OSI-774, CI-1033, EKB-569, GW-2016, E1.1,E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3; and7H-pyrrolo-[2,3-d]pyrimidine derivatives which are disclosed in WO03/013541.

Further anti-angiogenic compounds include compounds having anothermechanism for their activity, e.g., unrelated to protein or lipid kinaseinhibition, e.g., thalidomide (THALOMID®) and TNP-470.

Compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase are, e.g., inhibitors of phosphatase 1, phosphatase2A, PTEN or CDC_(25,) e.g., okadaic acid or a derivative thereof.

Compounds that induce cell differentiation processes are e.g. retinoicacid, α-, γ- or δ-tocopherol or α-, γ- or δ-tocotrienol.

The term cyclooxygenase inhibitor, as used herein, includes, but is notlimited to, e.g., Cox-2 inhibitors, 5-alkyl substituted2-arylaminophenylacetic acid and derivatives, such as celecoxib(CELEBREX®), rofecoxib (VIOXX®), etoricoxib, valdecoxib or a5-alkyl-2-arylaminophenylacetic acid, e.g.,5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid orlumiracoxib.

The term “bisphosphonates”, as used herein, includes, but is not limitedto, etridonic, clodronic, tiludronic, pamidronic, alendronic,ibandronic, risedronic and zoledronic acid. “Etridonic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark DIDRONEL®. “Clodronic acid” can be administered, e.g., in theform as it is marketed, e.g., under the trademark BONEFOS®. “Tiludronicacid” can be administered, e.g., in the form as it is marketed, e.g.,under the trademark SKELID®. “Pamidronic acid” can be administered,e.g., in the form as it is marketed, e.g., under the trademark AREDIA™.“Alendronic acid” can be administered, e.g., in the form as it ismarketed, e.g., under the trademark FOSAMAX®. “Ibandronic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark BONDRANAT®. “Risedronic acid” can be administered, e.g., inthe form as it is marketed, e.g., under the trademark ACTONEL®.“Zoledronic acid” can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ZOMETA®.

The term “mTOR inhibitors” relates to compounds which inhibit themammalian target of rapamycin (mTOR) and which possess antiproliferativeactivity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779and ABT578.

The term “heparanase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit heparin sulphate degradation. The termincludes, but is not limited to, PI-88.

The term “biological response modifier”, as used herein, refers to alymphokine or interferons, e.g., interferon γ.

The term “inhibitor of Ras oncogenic isoforms”, e.g., H-Ras, K-Ras orN-Ras, as used herein, refers to compounds which target, decrease orinhibit the oncogenic activity of Ras, e.g., a “farnesyl transferaseinhibitor”, e.g., L-744832, DK8G557 or R1 15777 (Zarnestra).

The term “telomerase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of telomerase. Compoundswhich target, decrease or inhibit the activity of telomerase areespecially compounds which inhibit the telomerase receptor, e.g.,telomestatin.

The term “methionine aminopeptidase inhibitor”, as used herein, refersto compounds which target, decrease or inhibit the activity ofmethionine aminopeptidase. Compounds which target, decrease or inhibitthe activity of methionine aminopeptidase are, e.g., bengamide or aderivative thereof.

The term “proteasome inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of the proteasome.Compounds which target, decrease or inhibit the activity of theproteasome include, e.g., PS-341 and MLN 341.

The term “matrix metalloproteinase inhibitor” or “MMP inhibitor”, asused herein, includes, but is not limited to, collagen peptidomimeticand nonpeptidomimetic inhibitors, tetracycline derivatives, e.g.,hydroxamate peptidomimetic inhibitor batimastat and its orallybioavailable analogue marimastat (BB-2516), prinomastat (AG3340),metastat (NSC 683551) BMS-279251, BAY 12-9566, TAA211, MMI270B orAAJ996.

The term “agents used in the treatment of hematologic malignancies”, asused herein, includes, but is not limited to, FMS-like tyrosine kinaseinhibitors, e.g., compounds targeting, decreasing or inhibiting theactivity of FMS-like tyrosine kinase receptors (Flt-3R); interferon,1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors,e.g., compounds which target, decrease or inhibit anaplastic lymphomakinase.

Compounds which target, decrease or inhibit the activity of FMS-liketyrosine kinase receptors (Flt-3R) are especially compounds, proteins orantibodies which inhibit members of the Flt-3R receptor kinase family,e.g., PKC412, midostaurin, a staurosporine derivative, SU1 1248 andMLN518.

The term “HSP90 inhibitors”, as used herein, includes, but is notlimited to, compounds targeting, decreasing or inhibiting the intrinsicATPase activity of HSP90; degrading, targeting, decreasing or inhibitingthe HSP90 client proteins via the ubiquitin proteasome pathway.Compounds targeting, decreasing or inhibiting the intrinsic ATPaseactivity of HSP90 are especially compounds, proteins or antibodies whichinhibit the ATPase activity of HSP90, e.g., 17-allylamino,17-demethoxygeldanamycin (17AAG), a geldanamycin derivative, othergeldanamycin related compounds, radicicol and HDAC inhibitors.

The term “antiproliferative antibodies”, as used herein, includes, butis not limited to, trastuzumab (HERCEPTIN™), Trastuzumab-DM1, erlotinib(TARCEVA™), bevacizumab (AVASTIN™), rituximab (RITUXAN®), PR064553(anti-CD40) and 2C₄ antibody. By antibodies is meant, e.g., intactmonoclonal antibodies, polyclonal antibodies, multispecific antibodiesformed from at least two intact antibodies, and antibodies fragments solong as they exhibit the desired biological activity. For the treatmentof acute myeloid leukemia (AML), compounds disclosed herein can be usedin combination with standard leukemia therapies, especially incombination with therapies used for the treatment of AML. In particular,compounds disclosed herein can be administered in combination with,e.g., farnesyl transferase inhibitors and/or other drugs useful for thetreatment of AML, such as Daunorubicin, Adriamycin, Ara-C, VP-16,Teniposide, Mitoxantrone, Idarubicin, Carboplatinum and PKC412.

A compound disclosed herein may also be used to advantage in combinationwith each other or in combination with other therapeutic agents,especially other anti-malarial agents. Such anti-malarial agentsinclude, but are not limited to proguanil, chlorproguanil, trimethoprim,chloroquine, mefloquine, lumefantrine, atovaquone,pyrimethamine-sulfadoxine, pyrimethamine-dapsone, halofantrine, quinine,quinidine, amodiaquine, amopyroquine, sulphonamides, artemisinin,arteflene, artemether, artesunate, primaquine, inhaled NO, L-arginine,Dipropylenetri-amine NONOate (NO donor), Rosiglitzone (PPAR-γ agonist),activated charcoal, Erythropoietin, Levamisole, and pyronaridine.

A compound disclosed herein may also be used to advantage in combinationwith each other or in combination with other therapeutic agents, such asused for the treatment of Leishmaniosis, Trypanosomiasis, Toxoplasmosisand Neurocysticercosis. Such agents include, but are not limited tochloroquine sulfate, atovaquone-proguanil, artemether-lumefantrine,quinine-sulfate, artesunate, quinine, doxycycline, clindamycin,meglumine antimoniate, sodium stibogluconate, miltefosine, ketoconazole,pentamidine, amphotericin B (AmB), liposomal-AmB, paromomycine,eflornithine, nifurtimox, suramin, melarsoprol, prednisolone,benznidazole, sulfadiazine, pyrimethamine, clindamycin, trimetropim,sulfamethoxazole, azitromycin, atovaquone, dexamethasone, praziquantel,albendazole, beta-lactams, fluoroquinolones, macrolides,aminoglycosides, sulfadiazine and pyrimethamine.

The structure of the active agents identified by code nos., generic ortrade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g., PatentsInternational, e.g., IMS World Publications.

The above-mentioned compounds, which can be used in combination with acompound disclosed herein, can be prepared and administered as describedin the art, such as in the documents cited above.

A compound disclosed herein may also be used to advantage in combinationwith known therapeutic processes, e.g., the administration of hormonesor especially radiation. A compound disclosed herein may in particularbe used as a radiosensitizer, especially for the treatment of tumorswhich exhibit poor sensitivity to radiotherapy.

By “combination”, there is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound disclosed herein and a combination partner may beadministered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g., synergistic, effect or any combination thereof. Theterms “coadministration” or “combined administration” or the like asutilized herein are meant to encompass administration of the selectedcombination partner to a single subject in need thereof (e.g. apatient), and are intended to include treatment regimens in which theagents are not necessarily administered by the same route ofadministration or at the same time. The term “pharmaceuticalcombination” as used herein means a product that results from the mixingor combining of more than one active ingredient and includes both fixedand non-fixed combinations of the active ingredients. The term “fixedcombination” means that the active ingredients, e.g. a compounddisclosed herein and a combination partner, are both administered to apatient simultaneously in the form of a single entity or dosage. Theterm “non-fixed combination” means that the active ingredients, e.g. acompound disclosed herein and a combination partner, are bothadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific time limits, wherein suchadministration provides therapeutically effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more activeingredients.

Methods of Treatment

In one embodiment, the methods of treatment disclosed herein compriseadministering a safe and effective amount of a compound or apharmaceutically composition disclosed herein to a patient in needthereof. Individual embodiments disclosed herein include methods oftreating any one of the above-mentioned disorders by administering asafe and effective amount of a compound disclosed herein or apharmaceutical composition containing a compound disclosed herein to apatient in need thereof.

In one embodiment, the compounds disclosed herein or pharmaceuticallycompositions containing the compounds disclosed herein may beadministered by any suitable route of administration, including bothsystemic administration and topical administration. Systemicadministration includes oral administration, parenteral administration,transdermal administration and rectal administration. Parenteraladministration is typically by injection or infusion, includingintravenous, intramuscular, and subcutaneous injection or infusion.Topical administration includes application to the skin as well asintraocular, otic, intravaginal, inhaled and intranasal administration.In one embodiment, the compounds disclosed herein or pharmaceuticalcompositions containing the compounds disclosed herein may beadministered orally. In another embodiment, the compounds disclosedherein or pharmaceutically compositions containing the compoundsdisclosed herein may be administered by inhalation. In a furtherembodiment, the compounds disclosed herein or pharmaceuticalcompositions containing the compounds disclosed herein may beadministered intranasally.

In another embodiment, the compounds disclosed herein orpharmaceutically compositions containing the compounds disclosed hereinmay be administered once or according to a dosing regimen wherein anumber of doses are administered at varying intervals of time for agiven period of time. For example, doses may be administered one, two,three, or four times per day. In one embodiment, a dose is administeredonce per day. In a further embodiment, a dose is administered twice perday. Doses may be administered until the desired therapeutic effect isachieved or indefinitely to maintain the desired therapeutic effect.Suitable dosing regimens for a compound disclosed herein or apharmaceutical composition containing a compound disclosed herein dependon the pharmacokinetic properties of that compound, such as absorption,distribution, and half-life, which can be determined by the skilledartisan. In addition, suitable dosing regimens, including the durationsuch regimens are administered, for a compound disclosed herein or apharmaceutical composition containing a compound disclosed herein dependon the disorder being treated, the severity of the disorder beingtreated, the age and physical condition of the patient being treated,the medical history of the patient to be treated, the nature ofconcurrent therapy, the desired therapeutic effect, and like factorswithin the knowledge and expertise of the skilled artisan. It will befurther understood by such skilled artisans that suitable dosingregimens may require adjustment given an individual patient's responseto the dosing regimen or over time as individual patient needs change.

The compound of the present invention may be administered eithersimultaneously with, or before or after, one or more other therapeuticagent. The compound of the present invention may be administeredseparately, by the same or different route of administration, ortogether in the same pharmaceutical composition as the other agents.

The pharmaceutical composition or combination of the present inventioncan be in unit dosage of about 1-1000 mg of active ingredient(s) for asubject of about 50-70 kg, or about 1-500 mg or about 1-250 mg or about1-150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredients.The therapeutically effective dosage of a compound, the pharmaceuticalcomposition, or the combinations thereof, is dependent on the species ofthe subject, the body weight, age and individual condition, the disorderor disease or the severity thereof being treated. A physician, clinicianor veterinarian of ordinary skill can readily determine the effectiveamount of each of the active ingredients necessary to prevent, treat orinhibit the progress of the disorder or disease. The above-cited dosageproperties are demonstrable in vitro and in vivo tests usingadvantageously mammals, e.g., mice, rats, dogs, monkeys or isolatedorgans, tissues and preparations thereof. The compounds of the presentinvention can be applied in vitro in the form of solutions, e.g.,aqueous solutions, and in vivo either enterally, parenterally,advantageously intravenously, e.g., as a suspension or in aqueoussolution.

In one embodiment, the therapeutically effective dose is from about 0.1mg to about 2,000 mg per day of a compound provided herein. Thepharmaceutical compositions therefore should provide a dosage of fromabout 0.1 mg to about 2000 mg of the compound. In certain embodiments,pharmaceutical dosage unit forms are prepared to provide from about 1 mgto about 2000 mg, from about 10 mg to about 1000 mg, from about 20 mg toabout 500 mg or from about 25 mg to about 250 mg of the essential activeingredient or a combination of essential ingredients per dosage unitform. In certain embodiments, the pharmaceutical dosage unit forms areprepared to provide about 10 mg, 20 mg, 25 mg, 50 mg, 100 mg, 250 mg,500 mg, 1000 mg or 2000 mg of the essential active ingredient.

Additionally, the compounds disclosed herein may be administered asprodrugs. As used herein, a “prodrug” of a compound disclosed herein isa functional derivative of the compound which, upon administration to apatient, eventually liberates the compound disclosed herein in vivo.Administration of a compound disclosed herein as a prodrug may enablethe skilled artisan to do one or more of the following: (a) modify theonset of the activity of the compound in vivo; (b) modify the durationof action of the compound in vivo; (c) modify the transportation ordistribution of the compound in vivo; (d) modify the solubility of thecompound in vivo; and (e) overcome a side effect or other difficultyencountered with the compound. Typical functional derivatives used toprepare prodrugs include modifications of the compound that arechemically or enzymatically cleavable in vivo. Such modifications, whichinclude the preparation of phosphates, amides, esters, thioesters,carbonates, and carbamates, are well known to those skilled in the art.

General Synthetic Procedures

In order to illustrate the invention, the following examples areincluded. However, it is to be understood that these examples do notlimit the invention and are only meant to suggest a method of practicingthe invention.

Generally, the compounds in this invention may be prepared by methodsdescribed herein, wherein the substituents are as defined for Formula(I), above, except where further noted. The following non-limitingschemes and examples are presented to further exemplify the invention.Persons skilled in the art will recognize that the chemical reactionsdescribed herein may be readily adapted to prepare a number of othercompounds of the invention, and alternative methods for preparing thecompounds of this invention are deemed to be within the scope of thisinvention. For example, the synthesis of non-exemplified compoundsaccording to the invention may be successfully performed bymodifications apparent to those skilled in the art, e.g., byappropriately protecting interfering groups, by utilizing other suitablereagents known in the art other than those described, and/or by makingroutine modifications of reaction conditions. Alternatively, otherreactions disclosed herein or known in the art will be recognized ashaving applicability for preparing other compounds of the invention.

In the examples described below, unless otherwise indicated alltemperatures are set forth in degrees Celsius. Reagents were purchasedfrom commercial suppliers such as Aldrich Chemical Company, ArcoChemical Company and Alfa Chemical Company, Shanghai Medpep. Co Ltd,Aladdin-Shanghai Jinchun Reagents, Ltd, and were used without furtherpurification unless otherwise indicated. Common solvents were purchasedfrom commercial suppliers such as Shantou XiLong Chemical Factory,Guangdong Guanghua Reagent Chemical Factory Co. Ltd., Guangzhou ReagentChemical Factory, Tainjin YuYu Fine Chemical Ltd., Qingdao TenglongReagent Chemical Ltd., and Qingdao Ocean Chemical Factory.

Anhydrous THF, dioxane, toluene, and ether were obtained by refluxingthe solvent with sodium. Anhydrous CH₂Cl₂ and CHCl₃ were obtained byrefluxing the solvent with CaH₂. EtOAc, PE, hexanes, DMA and DMF weretreated with anhydrous Na₂SO₄ prior use.

The reactions set forth below were done generally under a positivepressure of nitrogen or argon or with a drying tube (unless otherwisestated) in anhydrous solvents, and the reaction flasks were typicallyfitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried.

Column chromatography was conducted using a silica gel column. Silicagel (300-400 mesh) was purchased from Qingdao Ocean Chemical Factory.

¹H NMR spectra were recorded with a Bruker 400 MHz or 600 MHzspectrometer at ambient temperature. ¹H NMR spectra were obtained asCDCl₃, DMSO-d₆, CD₃OD or acetone-d₆ solutions (reported in ppm), usingTMS (0 ppm) or chloroform (7.26 ppm) as the reference standard. Whenpeak multiplicities are reported, the following abbreviations are used:s (singlet), d (doublet), t (triplet), m (multiplet), br (broadened), dd(doublet of doublets), dt (doublet of triplets), td (triplet ofdoublets). Coupling constants, when given, are reported in Hertz (Hz).

Low-resolution mass spectral (MS) data were generally determined on anAgilent 6120 quadrupole HPLC-MS (Zorbax SB-C18, 2.1×30 mm, 3.5 micron, 6minutes run, 0.6 mL/min flow rate, 5% to 95% (0.1% formic acid in CH₃CN)in (0.1% formic acid in H₂O)) with UV detection at 210/254 nm andelectrospray ionization (ESI).

Purities of compounds were assessed by Agilent 1260 pre-HPLC or Caleseppump 250 pre-HPLC (column: NOVASEP 50/80 mm DAC) with UV detection at210 nm and 254 nm.

The following abbreviations are used throughout the specification:

-   AcOH, HAc, CH₃COOH acetic acid-   Ac₂O acetic anhydride-   BnBr benzyl bromide-   BOC, Boc butyloxycarbony-   (Boc)₂O di-tert-butyl dicarbonate-   BH₃DMS borane-methyl sulfide complex-   BINAP 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl-   n-BuOH n-butyl alcohol-   CH₂Cl₂, DCM methylene chloride-   CDCl₃ chloroform deuterated-   CH₃I iodomethane-   DIEA, DIPEA, i-Pr₂NEt N,N-diisopropylethylamine-   DMF dimethylformamide-   DMP dimethyl phthalate-   DMAP 4-dimethylaminopyridine-   DMSO dimethylsulfoxide-   DHP dihydropyran-   PPTs pyridinium toluene-4-sulphonate-   Et₃N, TEA triethylamine-   EtOAc, EA ethyl acetate-   EtOH ethanol-   Et₂O diethyl ether-   EDCI N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride-   g gram-   h hour-   HATU 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate-   HCl hydrochloric acid-   HOAT 1-hydroxy-7-azabenzotriazole-   KOH potassium hydroxide-   KMnO₄ potassium permanganate-   K₂CO₃ potassium carbonate-   LiCl lithium chloride-   LiHMDS, LHMDS lithium bis(trimethylsilyl)amide-   LAH lithium aluminium hydride-   MeCN, CH₃CN acetonitrile-   MsCl methanesulfonyl chloride-   (NH₄)₂SO₄ ammonium sulfate-   NH₄Cl ammonium chloride-   NaH sodium hydride-   NaBH₃CN sodium cyanoborohydride-   Na₂CO₃ sodium carbonate-   NaHCO₃ sodium bicarbonate-   NaOH sodium hydroxide-   Na₂SO₄ sodium sulfate-   Na₂S₂O₃ sodium thiosulfate-   NaOAc ammonium acetate-   NBS bromosuccinimide-   MeOH methanol-   mL, ml milliliter-   Pd/C palladium on carbon-   PTSA p-toluenesulfonic acid-   PE petroleum ether (60-90° C.)-   PDC pyridinium dichromate-   RT, rt, r.t. room temperature-   PTSA p-toluenesulfonamide-   Pd(OAc)₂ palladium diacetate-   Pd/C palladium on activated carbon-   PDC pyridinium dichromate-   Rt retention time-   THF tetrahydrofuran-   TBAF tetrabutylammonium fluoride-   TFAA trifluoroacetic anhydride-   TFA, CF₃COOH trifluoroacetic acid-   Ti(Oi-Pr)₄ titanium tetraisopropanolate-   TsCl tosyl chloride

Representative synthetic procedures for the preparation of the compoundsdisclosed herein is outlined below in following Scheme 1 to Scheme 2.Unless otherwise indicated, each of Z, Z¹, R¹, R² and R⁴ carry thedefinitions set forth above in connection with Formula (I); p is 0, 1,2, 3, or 4; q is 0, 1, 2 or 3; PG is a protecting group; W is an azolyl.

Some compounds having Formula (6) or Formula (7) can be prepared by ageneral method illustrated in Scheme 1 and described in details in theExamples. As showing in Scheme 1, optionally substituteddichloropyrimidine compound (1) is reacted with optionally substitutedheterocyclic compound (2) with an aid of a base, such as Et₃N, to giveoptionally substituted heteroaryl compound (2). Compound (3) is coupledwith optionally substituted aminoindazole (4) or a hydrochloride thereofin the presence of a base, such as DIPEA, Et₃N, or in the presence of asuitable Pd catalyst, such as Pd(OAc)₂, to afford the compound (5). Theprotecting group of compound (5) was removed under acidic conditions,such as trifluoroacetic acid, a solution of HCl in EtOAc, to give thedesired protein kinase inhibitor (6). Other desired protein kinaseinhibitor having Formula (2) is obtained by introducing varioussubstituents to the compound (6) under appropriate conditions.

Some compounds having Formula (10) or Formula (11) can be prepared by ageneral method illustrated in Scheme 2 and described in details in theExamples. As showing in Scheme 2, Compound (3) is coupled withoptionally substituted aminoazole compound (8) or a hydrochloridethereof in the presence of a base, such as DIPEA, Et₃N, or in thepresence of a acid, such as trifluoroacetic acid, a solution of HCl inEtOAc, or in the presence of a suitable Pd catalyst, such as Pd(OAc)₂,to afford the compound (9). The protecting group of compound (9) wasremoved under acidic conditions, such as trifluoroacetic acid, asolution of HCl in EtOAc, or with an aid of a hydrazine hydrate or anyother appropriate condition to give the desired protein kinase inhibitor(10). Other desired protein kinase inhibitor having Formula (11) isobtained by introducing various substituents to the compound (10) underappropriate conditions.

EXAMPLES Example 1N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-methyl-N⁴-(2-azaspiro[4.5]decan-8-yl)pyrimidine-2,4-diamine

Step 1) ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate

To a suspension of NaH (60% mineral oil suspension, 33.3 g, 832.38 mmol)in anhydrous THF (1 L) was added a solution of1,4-dioxaspiro[4.5]decan-8-one (100 g, 640.29 mmol) in anhydrous THF(500 mL) dropwise at 0° C. for 1 h and continued to stir for 1 h. Thentriethyl phosphonoacetate (203.23 g, 832.38 mmol) was added to the abovesuspension dropwise at −20° C. in 1 h. The resulting mixture was allowedto warm to rt, stirred for 2 h, quenched with H₂O (1 L) and extractedwith EtOAC (1 L×3). The combined organic phases were washed with brine(1 L), dried over anhydrous Na₂SO₄, then filtered and concentrated invacuo. The residue was purified by silica gel column chromatography(EtOAc/PE (v/v)=1/10) to give the title compound as pale yellow oil (157g, 100%).

¹H NMR (600 MHz, CDCl₃): δ (ppm) 5.64 (s, 1H), 4.12 (q, J=7.1 Hz, 2H),3.95 (s, 4H), 2.97 (m, 2H), 2.36 (m, 2H), 1.74 (m, 4H), 1.25 (t, J=7.2Hz, 4H).

Step 2) ethyl 2-(8-(nitromethyl)-1,4-dioxaspiro[4.5]decan-8-yl)acetate

To a solution of ethyl 2-(1,4-dioxaspiro[4.5]decan-8-ylidene)acetate(157 g, 693.86 mmol) in THF (800 mL) was added nitromethane (55.8 mL,1.04 mmol) at rt followed by 1M solution of TBAF in THF (763.25 mL,763.25 mmol) which was added dropwise at rt. The reaction mixture washeated to reflux for 20 h and then concentrated in vacuo. The residuewas diluted with H₂O (800 mL) and extracted with EtOAc (800 mL×3). Thecombined organic phases were washed with brine (1 L), dried overanhydrous Na₂SO₄, then filtered and concentrated in vacuo. The residuewas purified by silica gel column chromatography (EtOAc/PE (v/v)=1/10)to give the title compound as pale yellow oil (165 g, 82.9%).

¹H NMR (600 MHz, CDCl₃): δ (ppm) 4.66 (s, 2H), 4.09 (q, J=7.1 Hz, 2H),3.88 (s, 4H), 2.49 (s, 2H), 1.64 (m, 8H), 1.21 (t, J=7.2 Hz, 4H).

Step 3) 1,4-dioxa-10-aza-dispiro[4.2.4.2]tetradecan-9-one

To a solution of ethyl2-(8-(nitromethyl)-1,4-dioxaspiro[4.5]decan-8-yl)acetate (109 g, 379.38mmol) in EtOH (600 mL) was added Raney Ni (10 g). The reaction mixturewas stirred under H₂ atmosphere at rt for 72 h and filtered. The filtercake was washed with a mixture of DCM and MeOH (1/1 (v/v), 500 mL). Thefiltrated was concentrated in vacuo and the residue was washed with amixture of PE and EtOAc (10/1 (v/v), 800 mL) and filtered to give theproduct as a white solid (67 g, 83.8%).

MS (ESI, pos. ion) m/z: 212.1 [M+H]⁺;

¹H NMR (600 MHz, CDCl₃): δ (ppm) 6.60 (s, 1H), 3.95 (s, 4H), 3.20 (s,2H), 2.23 (s, 2H), 1.72 (m, 4H), 1.65 (m, 4H).

Step 4) 1,4-dioxa-10-aza-dispiro[4.2.4.2]tetradecane

To a suspension of LiAlH₄ (21 g, 558.56 mmol) in anhydrous THF (500 mL)was added a solution of1,4-dioxa-10-aza-dispiro[4.2.4.2]tetradecan-9-one (59 g, 279.28 mmol) inanhydrous THF (400 mL) dropwise at 0° C. in 1 h. The reaction mixturewas heated to reflux and stirred for 2 h, then quenched with water at 0°C. until no bubble escaped and filtered. The filtere cake was washedwith a mixture of DCM and MeOH (1/1 (v/v), 500 mL). The filterate wasconcentrated in vacuo and the residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/10) to give the product as a yellowsolid (52 g, 94%).

MS (ESI, pos. ion) m/z: 198.2 [M+H]⁺;

¹H NMR (600 MHz, CDCl₃): δ (ppm) 3.88 (s, 4H), 2.90 (t, J=7.1 Hz, 2H),2.65 (s, 3H), 1.56 (ddd, J=29.3, 13.4, 6.9 Hz, 10H).

Step 5) 2-azaspiro[4.5]decan-8-one

To a solution of 1,4-dioxa-10-aza-dispiro[4.2.4.2]tetradecane (10 g,50.69 mmol) in MeOH (100 mL) was added HCl aqueous solution (2 M, 30 mL,60 mmol). The reaction mixture was stirred at rt overnight andconcentrated in vacuo. The residue was dissolved in water (100 mL) andadjusted to pH=10 with a saturated Na₂CO₃ aqueous solution, thenextrated with DCM (200 mL×3). The combined organic phases were washedwith brine (200 mL), dried over anhydrous Na₂SO₄, then filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/7) to give the title compound as ayellow solid (10 g, 100%).

MS (ESI, pos. ion) m/z: 154.2 [M+H]⁺.

Step 6) tert-butyl 8-oxo-2-azaspiro[4.5]decane-2-carboxylate

To a solution of 2-azaspiro[4.5]decan-8-one (10 g, 65.26 mmol) in amixture of THF and H₂O (3/1 (v/v), 200 mL) were added (Boc)₂O (28.48 g,130.53 mmol) and Na₂CO₃ (13.83 g, 130.53 mmol). The reaction mixture wasstirred at rt for 2 h, then quenched with H₂O (200 mL) and extractedwith EtOAc (300 mL×3). The combined organic phases were washed withbrine (500 mL), dried over Na₂SO₄, concentrated in vacuo. The residuewas purified by silica gel column chromatography (EtOAc/PE (v/v)=1/10)to give the product as colorless oil (9 g, 54.5%).

MS (ESI, pos. ion) m/z: 198.1 [(M-C₄H₈)+H]⁺.

Step 7) tert-butyl 8-(methylamino)-2-azaspiro[4.5]decane-2-carboxylate

To a solution of methylamine (33% [w/w] in EtOH, 7.42 g, 79.0 mmol) wasadded tert-butyl 8-oxo-2-azaspiro[4.5]decane-2-carboxylate (4 g, 15.80mmol), the reaction mixture was stirred at rt overnight. To the mixturewas added NaBH₃CN (2.98 g, 47.4 mmol), and the reaction mixture wasstirred at rt for another 2 h and concentrated in vacuo. The residue wasdiluted with H₂O (50 mL) and the resulting mixture was extracted withDCM (80 mL×3). The combined organic phases were washed with brine (100mL), dried over Na₂SO₄, filtered and concentrated in vacuo. The residuewas purified by silica gel column chromatography (DCM/MeOH (v/v)=10/1)to give the product as yellow oil (3.2g, 75.7%).

MS (ESI, pos. ion) m/z: 269.2 [M+H]⁺.

Step 8) tert-butyl8-((2-chloropyrimidin-4-yl)(methyl)amino)-2-azaspiro[4.5]decane-2-carboxylate

To a solution of 2,4-dichloropyrimidine (596 mg, 4.0 mmol) andtert-butyl 8-(methylamino)-2-azaspiro[4.5]decane-2-carboxylate (1.29 g,4.8 mmol) in EtOH (10 mL) was added Et₃N (0.92 g, 9.1 mmol) and thereaction mixture was stirred at rt overnight and then concentrated invacuo. The residue was purified by silica gel column chromatography(EtOAc/PE (v/v)=1/2) to give the product as a pale yellow solid (580 mg,38.1%).

MS (ESI, pos. ion) m/z: 381.2 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.01 (d, J=6.2 Hz, 1H), 6.31 (d, J=5.3Hz, 1H), 3.41 (ddd, J=20.4, 14.0, 7.1 Hz, 2H), 3.13 (d, J=25.2 Hz, 2H),2.93 (s, 2H), 1.81 (m, 2H), 1.65 (m, 8H), 1.48 (m, 11H).

Step 9) tert-butyl8-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)(methyl)amino)-2-azaspiro[4.5]decane-2-carboxylate

To a suspension of tert-butyl8-((2-chloropyrimidin-4-yl)(methyl)amino)-2-azaspiro[4.5]decane-2-carboxylate(500.0 mg, 1.3 mmol) and 2,3-dimethyl-2H-indazol-6-amine hydrochloride(778.6 mg, 3.9 mmol) in n-BuOH (10 mL) was added DIPEA (1.04 g, 8.0mmol). The reaction mixture was stirred at 150° C. in a sealed tubeovernight, then cooled down to rt, and concentrated in vacuo. Theresidue was purified by silica gel column chromatography (MeOH/DCM(v/v)=1/10) to give the title compound as a yellow solid (657 mg, 100%).

MS (ESI, pos. ion) m/z: 506.3 [M+H]⁺.

Step 10)N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-methyl-N⁴-(2-azaspiro[4.5]decan-8-yl)pyrimidine-2,4-diamine

To a solution of tert-butyl8-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)(methyl)amino)-2-azaspiro[4.5]decane-2-carboxylate(657 mg, 1.3 mmol) in DCM (10 mL) was added a solution of HCl in EtOAc(10 mL, 40 mmol). The reaction mixture was stirred at rt overnight andconcentrated in vacuo. The residue was dissolved in water (30 mL) andadjusted to pH=10 with a saturated Na₂CO₃ aqueous solution, thenextracted with DCM (250 mL×3). The combined organic phases were washedwith brine (250 mL), then dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/5) to give the title compound as abeige solid (0.30 g, 56.9%).

MS (ESI, pos. ion) m/z: 406.2 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 9.05 (s, 1H), 8.02 (s, 1H), 7.74 (d,J=6.7 Hz, 1H), 7.37 (d, J=8.9 Hz, 1H), 6.91 (d, J=8.8 Hz, 1H), 5.97 (d,J=6.6 Hz, 1H), 3.96 (s, 3H), 3.32 (m, 2H), 3.00 (s, 2H), 2.91 (s, 4H),2.49 (s, 3H), 1.93 (s, 1H), 1.87 (t, J=7.4 Hz, 3H), 1.71 (m, 6H).

Example 23-(8-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)(methyl)amino)-2-azaspiro[4.5]decan-2-yl)-3-oxopropanenitrile

To a solution ofN²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-methyl-N⁴-(2-azaspiro[4.5]decan-8-yl)pyrimidine-2,4-diamine(300 mg, 0.74 mmol), 2-cyanoacetic acid (156 mg, 1.83 mmol) in a mixtureof DCM and DMF (4/1 (v/v), 35 mL) were added HATU (925.8 mg, 2.44 mmol)and Et₃N (0.49 g, 4.84 mmol). After addition, the reaction mixture wasstirred at rt for 0.5 h, then quenched with H₂O (30 mL) and extractedwith DCM (100 mL×3). The combined organic phases were washed with brine(100 mL), and dried over anhydrous Na₂SO₄, then concentrated in vacuo.The residue was purified by silica gel column chromatography (DCM/MeOH(v/v)=20/1) to give the title compound as a beige solid (210 mg, 60.0%).

MS (ESI, pos. ion) m/z: 473.4 [M+H]⁺;

¹H NMR (600 MHz, CDCl₃): δ (ppm) 8.16 (d, J=9.2 Hz, 1H), 7.99 (dd,J=5.9, 3.3 Hz, 1H), 7.45 (dd, J=8.8, 4.4 Hz, 1H), 6.98 (m, 2H), 5.96 (t,J=5.5 Hz, 1H), 4.08 (d, J=9.3 Hz, 3H), 3.64 (dd, J=13.4, 6.7 Hz, 2H),3.44 (s, 3H), 3.36 (s, 1H), 2.96 (d, J=7.2 Hz, 3H), 2.59 (s, 3H), 2.02(t, J=7.1 Hz, 1H), 1.93 (t, J=7.3 Hz, 1H), 1.75 (m, 6H), 1.31 (s, 2H).

Example 35-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine

Step 1) tert-butyl 3a,4,7,7a-tetrahydro-1H-isoindole-2(3H)-carboxylate

To a suspension of LAH (22.80 g, 600 mmol) in THF (600 mL) at 0° C. wasadded tetrahydrophthalimide (39.45 g, 260.9 mmol) portionwise. Afteraddition, the reaction mixture was stirred at 60° C. for 18 h, thencooled down to 0° C. and quenched carefully with water (25 mL), followedby 15% KOH aqueous solutiuon (25 mL) and another 75 mL of water. Theresulting mixture was stirred at rt for 1 h and filtered through a padof Celite, then washed with DCM (500 mL). The filtrate was concentratedin vacuo to give isoindole as yellow oil, which was used in the nextstep without purification.

Accordingly, the isoindole in DCM (300 mL) was treated with Et₃N (39.61g, 391.4 mmol) and (Boc)₂O (68.32 g, 313.1 mmol) at 0° C. for 0.5 h andthen warmed to rt and stirred for another 21 h. The resulting mixturewas concentrated in vacuo and the residue was dissolved in EtOAc (600mL), then washed with citric acid aqueous solution (1 M, 2×130 mL),followed by saturated NaHCO₃ (2×130 mL) and brine (250 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄ then filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (EtOAc/PE (v/v)=1/5) to give the title compound as orangered oil (45.00 g, 77.3%).

MS (ESI, pos. ion) m/z: 168.2 [(M-C₄H₈)+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 5.64 (s, 2H), 3.40 (m, 2H), 3.16 (m,1H), 3.07 (m, 1H), 2.25 (m, 4H), 1.90 (m, 2H), 1.46 (s, 9H).

Step 2) 2,2′-(1-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)diacetic acid

To a solution of tert-butyl3a,4,7,7a-tetrahydro-1H-isoindole-2(3H)-carboxylate (4.91 g, 22 mmol)and (NH₄)₂SO₄ (1.55 g, 12 mmol) in H₂O (40 mL) was added KMnO₄ (8.20 g,52 mmol) portionwise at 5° C. in 0.5 h. The reaction mixture was stirredfor 6 h, then filtered and washed with H₂O (40 mL×3). The filtrate wasextracted with CH₂Cl₂ (40 mL×3) and the aqueous layer was adjusted topH=2-3 with 3 M HCl aqueous solution, then extracted with EtOAc (50mL×3). The combined EtOAc phases were washed with brine (50 mL×3), driedover Na₂SO₄, filtered and concentrated in vacuo to give the titlecompound as a pale yellow soild (4.52 g, 71.5%).

MS (ESI, pos. ion) m/z: 232.2 [(M-C₄H₈)+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 3.53 (m, 2H), 3.04 (m, 2H), 2.80 (m,2H), 2.44 (m, 4H), 1.43 (s, 9H).

Step 3) tert-butyl 5-oxohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a suspension of2,2′-(1-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)diacetic acid (3.40 g,11.8 mmol) in Ac₂O (21 mL) was added NaOAc (0.78 g, 9.5 mmol) and thereaction mixture was stirred at 120° C. for 3 h. After that, theresulting mixture was cooled down to rt then filtered and washed withEtOAc (20 mL×2). The filtrate was concentrated in vacuo and the residuewas purified by silica gel chromatography (EtOAc/PE (v/v)=1/4) to givethe title compound as orange yellow oil (1.38 g, 55.0%).

MS (ESI, pos. ion) m/z: 170.2 [(M-C₄H₈)+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 3.69 (m, 2H), 3.00 (m, 4H), 2.61 (dd,J=8.2, 18.4 Hz, 2H), 2.29 (dd, J=5.8, 18.4 Hz, 2H), 1.43 (s, 9H).

Step 4) tert-butyl5-(benzylamino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a solution of tert-butyl5-oxohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (9.57 g, 42.5 mmol)in DCM (170 mL) were added BnNH₂ (4.56 g, 42.5 mmol) and AcOH (2.55 g,42.5 mmol) at 0° C. and the reaction mixture was stirred at 0° C. for0.5 h. Then NaBH(OAc)₃ (18.00 g, 85.0 mmol) was added to the abovemixture and the resulting mixture was stirred at rt for another 20 h,then quenched with saturated NaHCO₃ solution (142 mL), and extractedwith DCM (250 mL×3). The combined organic phases were washed with brine(250 mL×3), dried over anhydrous Na₂SO₄, concentrated in vacuo. Theresidue was purified by silica gel column chromatography (EtOAc) to givethe title compound as a yellow solid (7.44 g, 55.3%).

MS (ESI, pos. ion) m/z: 317.3 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 7.31 (m, 4H), 7.26 (m, 1H), 3.79 (s,2H), 3.46 (m, 2H), 3.28 (d, J=8.8 Hz, 1H), 3.16 (tt, J=9.6, 6.9 Hz, 1H),2.55 (m, 2H), 2.28 (s, 2H), 2.22 (m, 2H), 1.45 (s, 9H), 1.31 (m, 2H).

Step 5) tert-butyl5-aminohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a solution of tert-butyl5-(benzylamino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (6.50 g,20.5 mmol) and AcOH (1.23 g, 20.5 mmol) in MeOH (150 mL) was addedPd(OH)₂/C (10% wt, 1.00 g), and the suspension was stirred at 40° C.under H₂ atmosphere overnight. The mixture was filtered through a pad ofCelite and then concentrated in vacuo. The residue was dissolved insaturated NaHCO₃ solution (70 mL) and extracted with DCM (100 mL×3). Thecombined organic phases were washed with brine (100 mL×3), dried overanhydrous Na₂SO₄, then filtered and concentrated in vacuo to give thetitle compound as a yellow solid (4.00 g, 86.2%).

MS (ESI, pos. ion) m/z: 227.2 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 3.44 (m, 2H), 3.31 (m, 3H), 2.98 (br.s, 2H), 2.57 (m, 2H), 2.22 (dt, J=14.0, 7.2 Hz, 2H), 1.44 (s, 9H).

Step 6) tert-butyl5-((2,5-dichloropyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a solution of 2,4,5-trichloropyrimidine (1.46 g, 7.96 mmol) andtert-butyl 5-aminohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (3.11g, 13.74 mmol) in EtOH (60 mL) was added Et₃N (2.21 g, 21.84 mmol) andthe reaction mixture was stirred at rt overnight and then concentratedin vacuo. The residue was dissolved in a mixture of EtOAc (50 mL) andwater (50 mL), and extracted with EtOAc (150 mL×3). The combined organicphases were washed with brine (150 mL), dried over anhydrous Na₂SO₄,then concentrated in vacuo. The residue was purified by silica gelcolumn chromatography (EtOAc/PE (v/v)=1/5) to give the title compound asa pale yellow solid (2.97 g, 100%).

MS (ESI, pos. ion) m/z: 373.0 [M+H]⁺.

Step 7) tert-butyl5-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a suspension of tert-butyl5-((2,5-dichloropyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate(621.8 mg, 1.666 mmol) and 2,3-dimethylindazol-6-amine hydrochloride(999.6 mg, 5.057 mmol) in n-BuOH (6 mL) was added DIPEA (1.10 g, 8.43mmol). The reaction mixture was stirred at 150° C. under microwave for 2h and concentrated in vacuo. The residue was purified by silica gelcolumn chromatography (EtOAc/PE (v/v)=2/1) to give the title compound asa pale brown solid (452.1 mg, 54.5%).

MS (ESI, pos. ion) m/z: 498.3 [M+H]⁺.

Step 8)5-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine

To a solution of tert-butyl5-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate(452.1 mg, 0.908 mmol) in DCM (10 mL) was added a solution of HCl inEtOAc (10 mL, 40 mmol). The reaction mixture was stirred at rt overnightand concentrated in vacuo. The residue was dissolved in water (30 mL)and adjusted to pH=10 with a saturated Na₂CO₃ aqueous solution, thenextracted with DCM (250 mL×3). The combined organic phases were washedwith brine (250 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/5) to give the title compound as abeige solid (179.6 mg, 49.7%).

MS (ESI, pos. ion) m/z: 398.3 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ (ppm) 9.14 (s, 1H), 8.10 (s, 1H), 7.96 (s,1H), 7.47 (d, J=8.9 Hz, 1H), 7.39 (d, J=6.4 Hz, 1H), 7.11 (d, J=8.9 Hz,1H), 4.45 (m, 1H), 3.96 (s, 3H), 3.17 (s, 1H), 3.05 (m, 2H), 2.92 (d,J=9.8 Hz, 2H), 2.73 (br. s, 2H), 2.53 (s, 3H), 2.30 (m, 2H), 1.492 (m,2H).

Example 43-(5-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-3-oxopropanenitrile

To a solution of5-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine(484.7 mg, 1.22 mmol) and 2-cyanoacetic acid (155.0 mg, 1.82 mmol) in amixture of DCM (40 mL) and DMF (10 mL) were added HATU (673.2 mg, 1.77mmol) and Et₃N (0.60 g, 5.90 mmol). The reaction mixture was stirred atrt for 5.5 h, then quenched with H₂O (30 mL) and extracted with DCM (100mL×3). The combined organic phases were washed with brine (100 mL),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. Theresidue was purified by silica gel column chromatography (DCM/MeOH(v/v)=40/1) to give the title compound as a beige solid (0.49 g, 87%).

MS (ESI, pos. ion) m/z: 465.0 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.09 (s, 1H), 7.94 (s, 1H), 7.47 (d,J=8.9 Hz, 1H), 6.98 (m, 2H), 5.27 (d, J=6.8 Hz, 1H), 4.53 (m, 1H), 4.07(s, 3H), 3.73 (m, 2H), 3.61 (dd, J=12.8, 4.3 Hz, 1H), 3.48 (dd, J=22.5,4.3 Hz, 1H), 3.42 (s, 2H), 2.90 (m, 2H), 2.64 (m, 2H), 2.60 (s, 3H),1.46 (m, 2H).

Example 5 5-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine

Step 1) tert-butyl 9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate

To a solution of tert-butyl 4-formylpiperidine-1-carboxylate (10.0 g,46.9 mmol) and KOH (1.3 g, 23.5 mmol) in EtOH (200 mL) was addedbut-3-en-2-one (3.9 g, 56.3 mmol), the mixture was stirred at 70° C. for16 h and concentrated in vacuo. The residue was purified by silica gelcolumn chromatography (EtOAc/PE (v/v)=1/4) to give the product as brownoil (5.2 g, 41.8%).

MS (ESI, pos. ion) m/z: 210.2 [M-55]⁺.

Step 2) tert-butyl 9-oxo-3-azaspiro[5.5]undecane-3-carboxylate

To a solution of tert-butyl9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate (5.2 g, 19.6 mmol) in DCM(80 mL) was added 10% Pd/C (0.5 g) and the suspention was stirred undera H₂ atmosphere at room temperature overnight. The reaction mixture wasfiltered and concentrated in vacuo, then the residue was purified bysilica gel column chromatography (EtOAc/PE (v/v)=1/4) to give theproduct as brown oil (3.1 g, 59.0%).

MS (ESI, pos. ion) m/z: 212.1 [M-55]⁺.

Step 3) tert-butyl 9-amino-3-azaspiro[5.5]undecane-3-carboxylate

To a solution of tert-butyl 9-oxo-3-azaspiro[5.5]undecane-3-carboxylate(5.35 g, 20.0 mmol) in EtOH (40 mL) were added a solution of NH₃ in MeOH(7M, 40 mL, 280.0 mmol) and Ti(Oi-Pr)₄ (11.30 g, 40.0 mmol), the mixturewas stirred at room temperature overnight. And then NaBH₄ (1.51 g, 40.0mmol) was added portionwise. After addition, the resulting mixture wasstirred at room temperature for another 5 h, then quenched with water(40 mL), stirred for 1 h and filtered. The filtrate was concentrated invacuo, and the residue was purified by silica gel column chromatography(MeOH/DCM (v/v)=1/25) to give the product as a light yellow solid (1.20g, 22.4%).

MS (ESI, pos. ion) m/z: 269.3 [M+H]⁺.

Step 4) tert-butyl9-((2,5-dichloropyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate

To a solution of 2,4,5-trichloropyrimidine (495.9 mg, 2.7 mmol) inethanol (20 mL) were added tert-butyl9-amino-3-azaspiro[5.5]undecane-3-carboxylate (1.08 g, 4.0 mmol) andEt₃N (413.6 mg, 4.1 mmol). The mixture was stirred at room temperaturefor 12 h and then concentrated in vacuo. The residue was purified bysilica gel column chromatography (EtOAc/PE (v/v)=1/10 to 1/7) to givethe product as a light yellow solid (387.1 mg, 34.5%).

MS (ESI, pos. ion) m/z: 415.0 [M+H]⁺.

Step 5) tert-butyl9-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate

To a suspension of tert-butyl9-((2,5-dichloropyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate(659.8 mg, 1.59 mmol) and 2,3-dimethylindazol-6-amine hydrochloride(580.6 mg, 2.94 mmol) in n-BuOH (6 mL) was added DIPEA (755.5 mg, 5.79mmol). The reaction mixture was stirred at 150° C. under microwaveradiation for 2 h and concentrated in vacuo. The residue was purified bysilica gel column chromatography (EtOAc/PE (v/v)=2/1) to give the titlecompound as a yellow solid (169.7 mg, 19.8%).

MS (ESI, pos. ion) m/z: 540.4 [M+H]⁺;

¹H NMR (600 MHz, CDCl₃): δ (ppm) 8.02 (d, J=1.2 Hz, 1H), 7.92 (s, 1H),7.45 (d, J=8.9 Hz, 1H), 7.07 (dd, J=8.9, 1.7 Hz, 1H), 7.00 (s, 1H), 5.17(d, J=7.4 Hz, 1H), 4.06 (s, 4H), 3.42 (m, 4H), 2.59 (s, 3H), 2.02 (d,J=9.5 Hz, 2H), 1.74 (m, 2H), 1.55 (br. s, 2H), 1.49 (s, 9H), 1.44 (m,6H).

Step 6)5-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine

To a solution of tert-butyl9-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate(161.5 mg, 0.30 mmol) in DCM (10 mL) was added a solution of HCl inEtOAc (10 mL, 40 mmol). The reaction mixture was stirred at rt for 2 hand concentrated in vacuo. The residue was dissolved in water (30 mL)and adjusted to pH=10 with a saturated Na₂CO₃ aqueous solution, thenextracted with DCM (100 mL×3). The combined organic phases were washedwith brine (100 mL), dried over anhydrous Na₂SO₄, concentrated in vacuo.The residue was purified by silica gel column chromatography (MeOH/DCM(v/v)=1/10) to give the title compound as a beige solid (131.6 mg,100%).

MS (ESI, pos. ion) m/z: 440.4 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 7.99 (s, 1H), 7.91 (s, 1H), 7.46 (d,J=8.9 Hz, 1H), 7.11 (dd, J=8.9, 1.5 Hz, 1H), 6.94 (s, 1H), 5.16 (d,J=7.4 Hz, 1H), 4.11 (t, J=6.7 Hz, 1H), 4.07 (s, 4H), 2.85 (m, 4H), 2.59(s, 3H), 2.00 (m, 2H), 1.78 (d, J=13.3 Hz, 2H), 1.56 (d, J=5.5 Hz, 2H),1.39 (m, 6H).

Example 63-(9-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)-3-azaspiro[5.5]undecan-3-yl)-3-oxopropanenitrile

To a solution of5-chloro-N²-(2,3-dimethyl-2H-indazol-6-yl)-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine(485.3 mg, 1.10 mmol) and 2-cyanoacetic acid (137.0 mg, 1.61 mmol) in amixture of DCM (40 mL) and DMF (10 mL) were added HATU (520.8 mg, 1.37mmol) and Et₃N (0.36 g, 3.55 mmol). After addition, the reaction mixturewas stirred at rt for 6 h, then quenched with H₂O (30 mL) and extractedwith DCM (100 mL×3). The combined organic phases were washed with brine(100 mL), dried over anhydrous Na₂SO₄, concentrated in vacuo. Theresidue was purified by silica gel column chromatography (MeOH/DCM(v/v)=1/50) to give the title compound as a beige solid (171.0 mg,30.6%).

MS (ESI, pos. ion) m/z: 507.3 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.06 (d, J=3.5 Hz, 1H), 7.93 (s, 1H),7.46 (d, J=8.9 Hz, 1H), 7.03 (m, 1H), 6.96 (s, 1H), 5.16 (d, J=6.8 Hz,1H), 4.10 (m, 1H), 4.06 (s, 3H), 3.63 (m, 2H), 3.49 (2, 2H), 3.45 (m,3H), 2.59 (s, 3H), 2.05 (m, 2H), 1.78 (m, 2H), 1.69 (m, 2H), 1.50 (m,6H).

Example 71-(9-((5-chloro-2-((2,3-dimethyl-2H-indazol-6-yl)amino)pyrimidin-4-yl)amino)-3-azaspiro[5.5]undecan-3-yl)ethanone

To a suspension ofN⁴-(9-azaspiro[5.5]undecan-3-yl)-5-chloro-N²-(2,3-dimethylindazol-6-yl)pyrimidine-2,4-diamine(0.10 g, 0.23 mmol) in anhydrous DCM (10.0 mL) were addedN,N-diethylethanamine (0.036 g, 0.35 mmol) and acetyl acetate (0.029 g,0.29 mmol). The mixture was stirred at room temperature for 30 min thendiluted with DCM (40 mL), washed with water (20 mL×2), followed bysaturated NaHCO₃ (20 mL) and brine (20 mL), then dried over anhydrousNa₂SO₄, filtered and concentrated in vacuo. The residue was purified bysilica gel column chromatography (DCM/MeOH (v/v)=30/1 to 15/1) to givethe title compound as a light green solid (75 mg, 68%).

MS (ESI, pos. ion) m/z: 482.0 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.03 (d, J=2.8 Hz, 1H), 7.90 (s, 1H),7.43 (d, J=8.9 Hz, 1H), 7.05-7.02 (m, 1H), 7.01 (s, 1H), 5.19-5.12 (m,1H), 4.08-4.00 (m, 4H), 3.62-3.54 (m, 2H), 3.45-3.38 (m, 2H), 2.57 (s,3H), 2.09 (s, 3H), 2.05-1.99 (m, 2H), 1.78-1.71 (m, 2H), 1.58-1.53 (m,2H), 1.51-1.43 (m, 6H).

Example 8N²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine

Step 1) tert-butyl5-((2-chloro-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a solution of 2,4-dichloro-5-methylpyrimidine (2.00 g, 12.3 mmol) andtert-butyl 5-aminohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (4.64g, 20.5 mmol) in EtOH (25 mL) was added Et₃N (2.62 g, 25.9 mmol). Thereaction mixture was stirred at 50° C. overnight and concentrated invacuo. The residue was purified by silica gel column chromatography(EtOAc/PE (v/v)=1/2) to give the title compound as yellow oil (2.42 g,55.9%).

MS (ESI, pos. ion) m/z: 353.0 [M+H]⁺;

¹H NMR (600 MHz, CDCl₃): δ (ppm) 7.68 (s, 1H), 5.25 (d, J=7.5 Hz, 1H),4.48 (m, 1H), 3.42 (m, 2H), 3.26 (d, J=11.1 Hz, 2H), 2.63 (m, 2H), 2.38(m, 2H), 1.92 (s, 3H), 1.41 (s, 9H), 1.33 (m, 2H).

Step 2) tert-butyl5-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate

To a suspension of tert-butyl5-((2-chloro-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate(421.5 mg, 1.19 mmol) and 2,3-dimethylindazol-6-amine hydrochloride(734.5 mg, 3.72 mmol) in n-BuOH (10 mL) was added DIPEA (795.2 mg, 6.09mmol). The reaction mixture was stirred at 150° C. under microwave for 2h and concentrated in vacuo. The residue was purified by silica gelcolumn chromatography (MeOH/DCM (v/v)=1/10) to give the title compoundas brown oil (570.5 mg, 100%).

MS (ESI, pos. ion) m/z: 478.2 [M+H]⁺.

Step 3)N²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine

To a solution of tert-butyl5-(2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxylatein DCM (15 mL) was added a solution of HCl in EtOAc (15 mL, 60 mmol).The reaction mixture was stirred at rt for 1 h and concentrated invacuo. The residue was dissolved in water (30 mL) and adjusted to pH=10with a saturated Na₂CO₃ aqueous solution, then extracted with DCM (250mL×3). The combined organic phases were washed with brine (250 mL),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. Theresidue was purified by silica gel column chromatography (MeOH/DCM(v/v)=1/5) to give the title compound as a beige solid (202.4 mg,42.8%).

MS (ESI, pos. ion) m/z: 378.3 [M+H]⁺;

¹H NMR (400 MHz, DMSO-d₆): δ (ppm) 9.20 (s, 1H), 9.13 (s, 1H), 7.96 (s,1H), 7.72 (s, 1H), 7.58 (d, J=8.8 Hz, 1H), 7.04 (d, J=8.8 Hz, 1H), 4.39(m, 1H), 4.00 (s, 3H), 3.06 (m, 2H), 2.81 (m, 2H), 2.57 (s, 3H), 2.37(m, 2H), 2.00 (s, 3H), 1.61 (m, 2H).

Example 93-(5-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-3-oxopropanenitrile

To a solution ofN²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine(86.9 mg, 0.23 mmol) and 2-cyanoacetic acid (31.3 mg, 0.37 mmol) in amixture of DCM (8 mL) and DMF (2 mL) were added HOAT (65.5 mg, 0.48mmol) and EDCI (77.6 mg, 0.40 mmol). After addition, the reactionmixture was stirred at 45° C. for 1 h, and then quenched with H₂O (30mL) and extracted with DCM (100 mL×3). The combined organic phases werewashed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/10) to give the title compound as abeige solid (69.8 mg, 68.2%).

MS (ESI, pos. ion) m/z: 445.0 [M+H]⁺.

¹H NMR (600 MHz, DMSO-d₆): δ (ppm) 8.90 (s, 1H), 8.14 (s, 1H), 7.69 (s,1H), 7.45 (d, J=8.9 Hz, 1H), 7.11 (d, J=8.9 Hz, 1H), 6.62 (s, 1H), 4.55(m, 1H), 3.96 (s, 3H), 3.58 (dd, J=10.4, 8.1 Hz, 2H), 3.51 (dd, J=12.1,8.5 Hz, 2H), 2.73 (m, 1H), 2.65 (m, 1H), 2.53 (s, 3H), 2.34 (m, 2H),1.93 (s, 3H), 1.49 (m, 2H).

Example 10N²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(2-(methylsulfonyl)octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine

Example 111-(5-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)ethanone

To a suspension ofN²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(octahydrocyclopenta[c]pyrrol-5-yl)pyrimidine-2,4-diamine(127.3 mg, 0.34 mmol) and N,N-diethylethanamine (68.3 mg, 0.68 mmol) inDCM (10 mL) was added methanesulfonyl chloride (64.2 mg, 0.56 mmol). Thereaction mixture was stirred at rt for 2 h, quenched with H₂O (30 mL)and extracted with DCM (100 mL×3). The combined organic phases werewashed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (MeOH/DCM (v/v)=1/20) to give example 10 as a beige solid(90.5 mg, 58.9%) and another elution (MeOH/DCM (v/v)=1/10) to giveexample 11 as a beige solid (50.2 mg, 35.5%).

Example 10

MS (ESI, pos. ion) m/z: 456.3 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.15 (s, 1H), 7.75 (s, 1H), 7.44 (d,J=8.9 Hz, 1H), 7.01 (dd, J=8.9, 1.5 Hz, 1H), 6.87 (br. s, 1H), 4.77 (d,J=7.5 Hz, 1H), 4.58 (m, 1H), 4.06 (s, 3H), 3.37 (s, 2H), 3.21 (dd,J=9.6, 7.0 Hz, 2H), 2.90 (m, 2H), 2.87 (s, 3H), 2.61 (m, 2H), 2.58 (s,3H), 1.99 (s, 3H), 1.51 (m, 2H).

Example 11

MS (ESI, pos. ion) m/z: 420.3 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃): δ (ppm) 8.13 (s, 1H), 7.76 (s, 1H), 7.44 (d,J=8.9 Hz, 1H), 7.02 (m, 1H), 6.86 (s, 1H), 4.59 (m, 1H), 4.50 (d, J=6.7Hz, 1H), 4.06 (s, 3H), 3.62 (m, 3H), 3.41 (dd, J=10.9, 3.4 Hz, 1H), 2.85(m, 2H), 2.66 (m, 2H), 2.59 (s, 3H), 2.08 (s, 3H), 1.96 (s, 3H), 1.41(m, 2H).

Example 123-(9-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecan-3-yl)-3-oxopropanenitrile

Step 1) tert-butyl9((2-chloro-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate

To a solution of 2,4-dichloro-5-methyl-pyrimidine (0.58 g, 3.60 mmol)and tert-butyl 9-amino-3-azaspiro[5.5]undecane-3-carboxylate (1.44 g,5.37 mmol) in EtOH (25 mL) was added Et₃N (0.86 g, 8.50 mmol). Afteraddition, the reaction mixture was stirred at 100° C. in a sealed tubefor 25.5 h and concentrated in vacuo. The residue was purified by silicagel column chromatography (EtOAc/PE (v/v)=1/2) to give the titlecompound as a yellow solid (332.2 mg, 24%).

MS (ESI, pos. ion) m/z: 395.4 [M+H]⁺.

Step 2) tert-butyl9-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate

A mixture of tert-butyl9-((2-chloro-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate(1.2 g, 3.0 mmol), 2,3-dimethylindazol-6-amine hydrochloride (896.5 mg,4.536 mmol), diacetoxypalladium (132.4 mg, 0.5898 mmol), BINAP (370.3mg, 0.5947 mmol) and dicesium carbonate (2.98 g, 9.15 mmol) wasdissolved in anhydrous dioxane (10 mL). The reaction mixture was stirredunder microwave irradiation at 150° C. for 2 h and then concentrated invacuo. The residue was purified by silica gel column chromatography(DCM/MeOH (v/v)=30/1) to give the title compound as a yellow solid (1.04g, 66%).

MS (ESI, pos. ion) m/z: 520.1 [M+H]⁺.

Step 3)N²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine

To tert-butyl9-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecane-3-carboxylate(1.04 g, 2.00 mmol) was added a solution of hydrogen chloride in ethylacetate (8 mL, 3 mol/L). The reaction was stirred at rt overnight andthen concentrated in vacuo. The residue was diluted with Na₂CO₃ solution(sat, 100 mL), the mixture was extracted with DCM/MeOH (10/1, 150 mL×3).The organic layers were dried with Na₂SO₄, filtered and concentrated invacuo to give the title compound as a yellow solid (804 mg, 95.8%).

MS (ESI, pos. ion) m/z: 420.0 [M+H]⁺.

Step 4)3-(9-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecan-3-yl)-3-oxopropanenitrile

To a solution ofN²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine(203.2 mg, 0.4843 mmol), HOAT (105.8 mg, 0.7773 mmol) and EDCI (144.5mg, 0.7538 mmol) in DCM/DMF (4 mL/1 mL) were added N,N-diethylethanamine(209.1 mg, 2.066 mmol) and 2-cyanoacetic acid (48.1 mg, 0.565 mmol). Thereaction was stirred at rt for 2.5 h and then concentrated in vacuo. Theresidue was purified by silica gel column chromatography (DCM/MeOH(v/v)=30/1) to give the title compound as a light pink solid (48.4 mg,20.4%).

MS (ESI, pos. ion) m/z: 487.3 [M+H]⁺;

¹H NMR (400 MHz, CDCl₃) δ (ppm): 8.52 (s, 1H), 8.11 (s, 1H), 7.60 (s,1H), 7.43 (d, J=8.9 Hz, 1H), 7.05 (d, J=7.3 Hz, 1H), 4.84 (dd, J=16.7,7.0 Hz, 1H), 4.11 (s, 1H), 4.03 (d, J=2.2 Hz, 3H), 3.66-3.53 (m, 2H),3.51-3.34 (m, 4H), 2.57 (s, 3H), 2.05 (d, J=10.9 Hz, 2H), 1.97 (s, 3H),1.75 (d, J=10.4 Hz, 2H), 1.68-1.41 (m, 8H).

Example 131-(9-((2-((2,3-dimethyl-2H-indazol-6-yl)amino)-5-methylpyrimidin-4-yl)amino)-3-azaspiro[5.5]undecan-3-yl)ethanone

To a solution ofN²-(2,3-dimethyl-2H-indazol-6-yl)-5-methyl-N⁴-(3-azaspiro[5.5]undecan-9-yl)pyrimidine-2,4-diamine(201.5 mg, 0.4802 mmol), HOAT (106.4 mg, 0.7817 mmol) and EDCI (145.7mg, 0.7600 mmol) in DCM/DMF (4 mL/lmL) were added N,N-diethylethanamine(207.6 mg, 2.052 mmol) and acetic acid (36.7 mg, 0.611 mmol). Thereaction was stirred at rt for 2 h. The reaction solution wasconcentrated in vacuo. The residue was purified by silica gel columnchromatography (DCM/MeOH (v/v)=30/1) to give the crude product as alight yellow solid (92 mg). The crude product was recrystallized fromacetone/MeOH (6 mL/0.5 mL) to give the title compound as a white solid(50.4 mg, 21.6%).

MS (ESI, pos. ion) m/z: 462.4[M+H]⁺.

Biological Testing

The LC/MS/MS system used in the analysis consists of an Agilent 1200Series vacuum degasser, binary pump, well-plate autosampler,thermostatted column compartment, the Agilent G6430 Triple QuadrupoleMass Spectrometer with an electrosprayionization (ESI) source.Quantitative analysis was carried out using MRM mode. The parameters forMRM transitions are in the Table A.

TABLE A MRM 490.2→383.1 Fragmentor 230 V CE 55 V Drying Gas Temp 350° C.Nebulize 40 psi Drying Gas Flow 10 L/min

An Agilent XDB-C18, 2.1×30 mm, 3.5 μM column was used for the analysis.5 μL of the samples were injected. Analysis condition: The mobile phasewas 0.1% formic acid in water (A) and 0.1% formic acid in methanol (B).The flow rate was 0.4 mL/min. And the gradient of Mobile phase was inthe Table B.

TABLE B Time Gradient of Mobile Phase B 0.5 min 5% 1.0 min 95% 2.2 min95% 2.3 min 5% 5.0 min stop

Alternatively, an Agilent 6330 series LC/MS/MS spectrometer equippedwith G1312A binary pumps, a G1367A autosampler and a G1314C UV detectorwere used in the analysis. An ESI source was used on the LC/MS/MSspectrometer. The analysis was done in positive ion mode as appropriateand the MRM transition for each analyte was optimized using standardsolution. A Capcell MP-C18 100×4.6 mm I.D., 5 μM column (Phenomenex,Torrance, Calif., USA) was used during the analysis. The mobile phasewas 5 mM ammonia acetate, 0.1% MeOH in water (A): 5 mM ammonia acetate,0.1% MeOH in acetonitrile (B) (70:30, v/v). The flow rate was 0.6mL/min. Column was maintained at ambient temperature. 20 μL of thesamples were injected.

Example A Compound Stability in Human and Rat Liver Microsomes

Human or rat liver microsomes incubations were conducted in duplicate inpolypropylene tubes. The typical incubation mixtures consisted of humanor rat liver microsomes (0.5 mg protein/mL), compounds of interest (5μM) and NADPH (1.0 mM) in a total volume of 200 μL potassium phosphatebuffer (PBS, 100 mM, pH 7.4). Compounds were dissolved in DMSO anddiluted with PBS such that the final concentration of DMSO was 0.05%.The enzymatic reactions were commenced with the addition of proteinafter a 3-min preincubation and incubated in a water bath open to theair at 37° C. Reactions were terminated at various time points (0, 5,10, 15, 30, 60 min) by adding equal volume of ice-cold acetonitrile. Thesamples were stored at 80° C. until LC/MS/MS assays.

The concentrations of compounds in the incubation mixtures of human orrat liver microsomes were determined by a LC/MS/MS method. The ranges ofthe linearity in the concentration range were determined for each testedcompounds.

A parallel incubation was performed using denatured microsomes as thenegative control, and reactions were terminated at various time points(0, 15, 60 min) after incubation at 37° C.

Dextromethorphan (70 μM) was selected as the positive control, andreactions were terminated at various time points (0, 5, 10, 15, 30, 60min) after incubation at 37° C. Both positive and negative controlsamples were included in each assay to ensure the integrity of themicrosomal incubation system.

Data Analysis

The concentrations of compounds in human or rat liver microsomeincubations were plotted as a percentage of the relevant zero time pointcontrol for each reaction. The in vivo CL_(int) were extrapolated (ref.:Naritomi, Y.; Terashita, S.; Kimura, S.; Suzuki, A.; Kagayama, A.; andSugiyama, Y.; Prediction of human hepatic clearance from in vivo animalexperiments and in vitro metabolic studies with liver microsomes fromanimals and humans. Drug Metab. Dispos., 2001, 29: 1316-1324).

Exemplary results from selected compounds of the invention are listed inTable 2. The compounds disclosed herein exhibited desirable stabilitywhen the compounds were incubated in human and rat liver microsomes.

TABLE 2 Stability of selected compounds of the invention in human andrat liver microsomes Human Rat T_(1/2) CL_(int) T_(1/2) CL_(int) Example# (min) (mL/min/kg) (min) (mL/min/kg) Ex. 2 20.34 85.46 22.19 111.93 Ex.4 14.30 121.56 25.26 98.33 Ex. 6 79.85 21.77 40.95 60.65 Ex. 7 42.940.52 29.3 84.77 Ex. 9 31.2 55.72 52.7 47.13 Ex. 10 19.99 86.96 50.7448.95 Ex. 12 166.4 10.45 57.78 42.99 Ex. 13 51.66 33.65 27.86 89.15

Example B Evaluation of Pharmacokinetics After Intravenous and OralAdministration of the Compounds Disclosed Herein in Mice, Rats, Dogs andMonkeys

The compounds disclosed herein are assessed in pharmacokinetic studiesin mice, rats, dogs or monkeys. The compounds are administered as awater solution, 2% HPMC+1% TWEEN®80 in water solution, 5% DMSO+5%solutol in saline, 4% MC suspension or capsule. For the intravenousadministration, the animals are generally given at 1 or 2 mg/kg dose.For the oral (p.o.) dosing, mice and rats are generally given 5 or 10mg/kg dose, and dogs and monkeys are generally given 10 mg/kg dose. Theblood samples (0.3 mL) are drawn at 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 6.0,8.0, 12 and 24 h time points or 0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0,8.0 and 24 h time points and centrifuged at 3,000 or 4000 rpm for 2 to10 min. The plasma solutions are collected, and stored at −20° C. or−70° C. until analyzed by LC/MS/MS as described above.

Exemplary study results from examples disclosed herein are listed inTable 3. The compounds disclosed herein exhibited optimizedpharmacokinetic properties with good absorption, desirable oralbioavailability (F) and half-life (T1/2) when the compounds wereadministered orally or intravenously.

TABLE 3 Pharmacokinetic profiles of selected compounds of the inventionin rats iv dosing dose T_(1/2) AUC_(last) Cl/F Vss F Example # (mg/kg)(h) (ng · h/ml) (L/h/kg) (L/kg) (%) Ex. 2 1 1.81 332 2.99 3.59 18.3 Ex.4 1 1.36 458 2.17 1.44 59.0 Ex. 6 1 1.31 743 1.34 2.55 52.8 Ex. 7 1 0.85688 1.47 1.68 66.0 Ex. 10 1 0.73 501 1.99 1.75 16.1 Ex. 12 1 0.90 1446.84 5.71 24.9 Ex. 13 1 0.64 238 4.53 3.50 22.1

Example C Kinase Activity Assay

The efficacy of the compounds disclosed herein as inhibitors of proteinkinases can be evaluated as follows.

General Description for Kinase Assays

Kinase assays can be performed by measurement of incorporation of γ-³³PATP into immobilized myelin basic protein (MBP). High binding white 384well plates (Greiner) are coated with MBP (Sigma #M-1891) by incubationof 60 μL/well of 20 μg/mL MBP in Tris-buffered saline (TBS; 50 mM TrispH 8.0, 138 mM NaCl, 2.7 mM KCl) for 24 h at 4° C. Plates are washed 3×with 100 μL TBS. Kinase reactions are carried out in a total volume of34 μL in kinase buffer (5 mM Hepes pH 7.6, 15 mM NaCl, 0.01% bovinegamma globulin (Sigma #I-5506), 10 mM MgCl₂, 1 mM DTT, 0.02%TritonX-100). Compound dilutions are performed in DMSO and added toassay wells to a final DMSO concentration of 1%. Each data point ismeasured in duplicate, and at least two duplicate assays are performedfor each individual compound determination. Enzyme is added to finalconcentrations of 10 nM or 20 nM, for example. A mixture of unlabeledATP and γ-³³P ATP is added to start the reaction (2×10⁶ cpm of γ-³³P ATPper well (3000 Ci/mmole) and 10 μM unlabeled ATP, typically. Thereactions are carried out for 1 h at room temperature with shakingPlates are washed 7× with TBS, followed by the addition of 50 μL/wellscintillation fluid (Wallac). Plates are read using a Wallac Triluxcounter. This is only one format of such assays; various other formatsare possible, as known to one skilled in the art.

The above assay procedure can be used to determine the ICso forinhibition and/or the inhibition constant, K. The IC₅₀ is defined as theconcentration of compound required to reduce the enzyme activity by 50%under the condition of the assay. The IC₅₀ value is estimated bypreparing a 10 point curve using a ½ log dilution series (for example, atypical curve may be prepared using the following compoundconcentrations: 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM,0.001 μM, 0.0003 μM and 0 μM).

Kianse General Assay Protocol JAK1 (h)

JAK1 (h) is incubated with 20 mM Tris/HCl pH 7.5, 0.2 mM EDTA, 500 μMGEEPLYWSFPAKKK, 10 mM MgAcetate and [γ-³³P-ATP] (specific activityapprox. 500 cpm/pmol, concentration as required). The reaction isinitiated by the addition of the MgATP mix. After incubation for 40minutes at room temperature, the reaction is stopped by the addition of3% phosphoric acid solution. 10 μL of the reaction is then spotted ontoa P30 filtermat and washed three times for 5 minutes in 75 mM phosphoricacid and once in methanol prior to drying and scintillation counting.

JAK2 (h)

JAK2 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 100 μMKTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC, 10 mM MgAcetate and [γ-³³P-ATP](specific activity approx. 500 cpm/pmol, concentration as required). Thereaction is initiated by the addition of the MgATP mix. After incubationfor 40 minutes at room temperature, the reaction is stopped by theaddition of 3% phosphoric acid solution. 10 μL of the reaction is thenspotted onto a P30 filtermat and washed three times for 5 minutes in 75mM phosphoric acid and once in methanol prior to drying andscintillation counting.

JAK3 (h)

JAK3 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 500 μMGGEEEEYFELVKKKK, 10 mM MgAcetate and [γ-³³P-ATP] (specific activityapprox. 500 cpm/pmol, concentration as required). The reaction isinitiated by the addition of the MgATP mix. After incubation for 40minutes at room temperature, the reaction is stopped by the addition of3% phosphoric acid solution. 10 μL of the reaction is then spotted ontoa P30 filtermat and washed three times for 5 minutes in 75 mM phosphoricacid and once in methanol prior to drying and scintillation counting.

TYK2 (h)

TYK2 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250 μMGGMEDIYFEFMGGKKK, 10 mM MgAcetate and [γ-³³P-ATP] (specific activityapprox. 500 cpm/pmol, concentration as required). The reaction isinitiated by the addition of the MgATP mix. After incubation for 40minutes at room temperature, the reaction is stopped by the addition of3% phosphoric acid solution. 10 μL of the reaction is then spotted ontoa P30 filtermat and washed three times for 5 minutes in 75 mM phosphoricacid and once in methanol prior to drying and scintillation counting.

FLT3 (h)

FLT3 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 50 μMEAIYAAPFAKKK, 10 mM MgAcetate and [γ-³³P-ATP] (specific activity approx.500 cpm/pmol, concentration as required). The reaction is initiated bythe addition of the MgATP mix. After incubation for 40 minutes at roomtemperature, the reaction is stopped by the addition of 3% phosphoricacid solution. 10 μL of the reaction is then spotted onto a P30filtermat and washed three times for 5 minutes in 75 mM phosphoric acidand once in methanol prior to drying and scintillation counting.

Aurora-A (h)

Aurora-A (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 200 μMLRRASLG (Kemptide), 10 mM MgAcetate and [γ-³³P-ATP] (specific activityapprox. 500 cpm/pmol, concentration as required). The reaction isinitiated by the addition of the MgATP mix. After incubation for 40minutes at room temperature, the reaction is stopped by the addition of3% phosphoric acid solution. 10 μL of the reaction is then spotted ontoa P30 filtermat and washed three times for 5 minutes in 75 mM phosphoricacid and once in methanol prior to drying and scintillation counting.

Aurora-B (h)

Aurora-B (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 30 μMAKRRRLSSLRA, 10 mM MgAcetate and [γ-³³PATP] (specific activity approx.500 cpm/pmol, concentration as required). The reaction is initiated bythe addition of the MgATP mix. After incubation for 40 minutes at roomtemperature, the reaction is stopped by the addition of a 3% phosphoricacid solution. 10 μL of the reaction is then spotted onto a P30filtermat and washed three times for 5 minutes in 75 mM phosphoric acidand once in methanol prior to drying and scintillation counting.

The kinase assays described herein were performed at Millipore UK Ltd,Dundee Technology Park, Dundee DD2 1SW, UK.

Alternatively, the kinase activities of the compounds can be measuredusing KINOMEscan™, which is based on a competition binding assay thatquantitatively measures the ability of a compound to compete with animmobilized, active-site directed ligand. The assay was performed bycombining three components: DNA-tagged kinase; immobilized ligand; and atest compound. The ability of the test compound to compete with theimmobilized ligand was measured via quantitative PCR of the DNA tag.

For most assays, kinase-tagged T7 phage strains were prepared in an E.coli host derived from the BL21 strain. E. coli were grown to log-phaseand infected with T7 phage and incubated with shaking at 32° C. untillysis. The lysates were centrifuged and filtered to remove cell debris.The remaining kinases were produced in HEK-293 cells and subsequentlytagged with DNA for qPCR detection. Streptavidin-coated magnetic beadswere treated with biotinylated small molecule ligands for 30 minutes atroom temperature to generate affinity resins for kinase assays. Theliganded beads were blocked with excess biotin and washed with blockingbuffer (SEABLOCK™ (Pierce), 1% BSA, 0.05% TWEEN®20, 1 mM DTT) to removeunbound ligand and to reduce nonspecific binding. Binding reactions wereassembled by combining kinases, liganded affinity beads, and testcompounds in 1× binding buffer (20% SEABLOCK™, 0.17× PBS, 0.05%TWEEN®20, 6 mM DTT). All reactions were performed in polystyrene 96-wellplates in a final volume of 0.135 mL. The assay plates were incubated atroom temperature with shaking for 1 hour and the affinity beads werewashed with wash buffer (1× PBS, 0.05% TWEEN 20). The beads were thenre-suspended in elution buffer (1× PBS, 0.05% TWEEN®20, 0.5 μMnon-biotinylated affinity ligand) and incubated at room temperature withshaking for 30 minutes. The kinase concentration in the eluates wasmeasured by qPCR.

The kinase activity assays described herein can be performed usingKINOMEscan™ Profiling Service at DiscoveRx Corporation, 42501 Albrae St.Fremont, Calif. 94538, USA.

Exemplary assay results from compounds disclosed herein are listed inTable 4 and Table 5. The compounds disclosed herein displayed potentinhibitory activities against JAK1, JAK2, JAK3, TYK2, Aurora-A and/orFLT3 kinases in the corresponding kinase assays.

Table 4 lised the IC₅₀s of some compounds described herein in the JAK1and JAK2 kinase assays, Table 5 lised the IC₅₀s of some compoundsdescribed herein in the JAK3, TYK2, Aurora-A and FLT3 kinase assays.

TABLE 4 JAK1 and JAK2 Kinase inhibition data IC₅₀ (nM) Example # JAK1JAK2 Ex. 4 1 9 Ex. 6 0.6 2 Ex. 7 0.9 2 Ex. 9 2 NT Ex. 10 7 NT Ex. 11 57NT Ex. 12 1 NT Ex. 13 2 NT NT: not tested

TABLE 5 JAK3, TYK2, Aurora-A and FLT3 Kinase inhibition data IC₅₀ (nM)Example # JAK3 TYK2 Aurora-A FLT3 Ex. 4 535 20 104 545 Ex. 7 142 NT NT 3NT: not tested

Finally, it should be noted that there are alternative ways ofimplementing the present invention. Accordingly, the present embodimentsare to be considered as illustrative and not restrictive and theinvention is not be limited to the details given herein, but may bemodified within the scope and equivalents of the appended claims. Allpublications and patents cited herein are incorporated by reference.

What is claimed is:
 1. A compound having Formula (I):

or a stereoisomer, a tautomer, an N-oxide, a solvate, a metabolite, apharmaceutically acceptable salt or a prodrug thereof, wherein: Z isC₇-C₁₂ spiro bicycloalkyl, C₇-C₁₂ fused bicycloalkyl, 7-12 memberedspiro heterobicyclyl or 7-12 membered fused heterobicycloalkyl, whereinZ is optionally substituted by 1, 2, 3, 4 or 5R² groups; Z¹ is H, C₁-C₁₂alkyl, C₃-C₁₂ cycloalkyl or 3-12 membered heterocyclyl, wherein Z¹ isoptionally substituted by 1, 2, 3, 4 or 5R³ groups; A is indazolyl orpyrazolopyridinyl,wherein A is optionally substituted by 1, 2, 3, 4 or5R⁴ groups; R¹ is H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂ alkyl, C₁-C₁₂haloalkyl, C₁-C₁₂ alkoxyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂cycloalkyl, 3-12 membered heterocyclyl, C₆-C₁₂ aryl, 5-12 memberedheteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein R¹ is optionally substituted by 1, 2, 3, 4 or 5R⁸ groups; eachR² is independently H, F, Cl, Br, I, NO₂, N₃, CN, OH, NH₂, 'C(═O)CH₂CN,C₁-C₁₂ alkyl, C₁-C₁₂ haloalkyl, C₁-C₁₂ alkoxy, C₂-C₁₂ alkenyl, C₂-C₁₂alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 membered heterocyclyl,5-12 membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b),—C(═O)R⁵, —S(═O)₂R⁵, —OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)—OR^(c),—N(R⁹C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c), —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—C(═NR^(c))NR^(a)R^(b), —N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵or —S(═O)₂NR^(a)R^(b), or two adjacent R² taken together with the atomsto which they are attached form a C₃-C₁₂ cycloalkyl or 3-12 memberedheterocycloalkyl group, wherein each of the above substituents isoptionally independently substituted by 1, 2, 3, 4 or 5R⁸ groups; eachR³ and R⁴ is independently H, F, Cl, Br, I, NO₂, N₃, CN, C₁-C₁₂ alkyl,C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, —(C₁-C₄alkylene)-(C₃-C₁₂ cycloalkyl), C₆-C₁₂ aryl, 3-12 membered heterocyclyl,—(C₁-C₄ alkylene)-(3-12 membered heterocyclyl), 5-12 memberedheteroaryl, —(CR⁶R⁷)_(n)OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—OC(═O)R⁵, —O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —C(═NR^(c))NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),wherein each R³ and R⁴ is optionally independently substituted by 1, 2,3, 4 or 5R⁸ groups; each R⁵ is independently H, C₁-C₁₂ alkyl, C₁-C₁₂haloalkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂aryl, 3-12 membered heterocyclyl or 5-12 membered heteroaryl, whereineach R⁵ is optionally independently substituted by 1, 2, 3, 4 or 5R⁸groups; each R⁶ and R⁷ is independently H, F, Cl, Br, I, NO₂, N₃, CN,C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂aryl, 3-12 membered heterocyclyl or 5-12 membered heteroaryl, or R⁶ andR⁷ taken together with the carbon atom to which they are attached form aC₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 membered heterocyclyl or 5-12membered heteroaryl group, wherein each of the above substituents isoptionally independently substituted by 1, 2, 3, 4 or 5R⁸ groups; eachR⁸ is independently F, Cl, Br, I, CN, NO₂, N₃, C₁-C₁₂ alkyl, C₂-C₁₂alkenyl, C₂-C₁₂ alkynyl, C₃-C₁₂ cycloalkyl, C₆-C₁₂ aryl, 3-12 memberedheterocyclyl, 5-12 membered heteroaryl, NH₂, —NH(C₁-C₁₂ alkyl),—NH(CH₂)_(n)—(C₃-C₁₂ cycloalkyl), —NH(CH₂)_(n)—(C₆-C₁₂ aryl),—NH(CH₂)_(n)-(3-12 membered heterocyclyl), —NH(CH₂)_(n)-(5-12 memberedheteroaryl), —N(C₁-C₁₂ alkyl)₂, —N[(CH₂)_(n)—(C₃-C₁₂ cycloalkyl)]₂,—N[(CH₂)_(n)—(C₆-C₁₂ aryl)]₂, —N[(CH₂)_(n)-(3-12 memberedheterocyclyl)]₂, —N[(CH₂)_(n)-(5-12 membered heteroaryl)]₂, OH,—O(C₁-C₁₂ alkyl), —O(CH₂)_(n)—(C₃-C₁₂cycloalkyl), —O(CH₂)_(n)—(C₆-C₁₂aryl), —O(CH₂)_(n)-(3-12 membered heterocyclyl) or —O(CH₂)_(n)-(5-12membered heteroaryl); each R^(a), R^(b) and R^(c) is independently H,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₄alkylene)-(C₃-C₆ cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₄alkylene)-(3-6 membered heterocyclyl), C₆-C₁₀ aryl, —(C₁-C₄alkylene)-(C₆-C₁₀ aryl), 5-10 membered heteroaryl or —(C₁-C₄alkylene)-(5-10 membered heteroaryl), or R^(a) and R^(b) taken togetherwith the nitrogen atom to which they are attached form a 3-8 memberedheterocyclyl group, wherein each of the above substituents is optionallyindependently substituted by 1, 2, 3 or 4 substitutents independentlyselected from F, Cl, Br, CN, N₃, OH, NH₂, C₁-C₆ alkyl, C₁-C₆ haloalkyl,C₁-C₆ alkoxy and C₁-C₆ alkylamino; each m is independently 1 or 2; andeach n is independently 0, 1, 2, 3 or
 4. 2. The compound of claim 1,wherein Z is C₈-C₁₁ spiro bicycloalkyl, C₈-C₁₀ fused bicycloalkyl, 8-11membered spiro heterobicyclyl or 8-10 membered fused heterobicycloalkyl,wherein Z is optionally substituted by 1, 2, 3 or 4R² groups.
 3. Thecompound of claim 1, wherein Z¹ is H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl or3-6 membered heterocyclyl, wherein Z¹ is optionally substituted by 1, 2or 3R³ groups.
 4. The compound of claim 1, wherein R¹ is H, F, Cl, CN,N₃, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl, C₁-C₆alkoxyl, C₃-C₆ cycloalkyl, 3-6 membered heterocyclyl,—(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or —S(═O)₂NR^(a)R^(b), wherein R¹ isoptionally substituted by 1, 2 or 3R⁸ groups.
 5. The compound of claim1, wherein each R² is independently H, F, Cl, CN, N₃, NO₂, OH, NH₂,—C(═O)CH₂CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl,C₁-C₆ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 membered heterocyclyl, 5-6membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)NR^(a)R^(b),—C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)OR^(c),—N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b),—N(R^(c))C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or —S(═O)₂NR^(a)R^(b),or two adjacent R² taken together with the atoms to which they areattached form a C₃-C₆ cycloalkyl, or 3-6 membered heterocycloalkylgroup, wherein each of the above substituents is optionallyindependently substituted by 1, 2 or 3R⁸ groups.
 6. The compound ofclaim 1, wherein each R³ and R⁴ is independently H, F, Cl, Br, I, NO₂,N₃, CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl,—(C₁-C₂ alkylene)-(C₃-C₆ cycloalkyl), phenyl, 3-6 membered heterocyclyl,—(C₁-C₂ alkylene)-(3-6 membered heterocyclyl), 5-6 membered heteroaryl,—(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵, —OC(═O)R⁵,—O(CR⁶R⁷)_(n)—R⁵, —N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)OR^(c),—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —N(R^(c))S(═O)_(m)R⁵ or—S(═O)₂NR^(a)R^(b), wherein each R³ and R⁴ is optionally independentlysubstituted by 1, 2 or 3R⁸ groups.
 7. The compound of claim 1, whereineach R⁵ is independently H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6 membered heterocyclyl or5-6 membered heteroaryl, wherein each R⁵ is optionally independentlysubstituted by 1, 2 or 3R⁸ groups.
 8. The compound of claim 1, whereineach R⁶ and R⁷ is independently H, F, Cl, Br, I, CN, N₃, NO₂, C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6membered heterocyclyl or 5-6 membered heteroaryl, or R⁶ and R⁷ takentogether with the carbon atom to which they are attached form a C₃-C₆cycloalkyl, phenyl, 3-6 membered heterocyclyl or 5-6 membered heteroarylgroup, wherein each of the above substituents is optionallyindependently substituted by 1, 2 or 3R⁸ groups.
 9. The compound ofclaim 1, wherein each R⁸ is independently F, Cl, CN, N₃, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6 memberedheterocyclyl, 5-6 membered heteroaryl, NH₂, —NH(C₁-C₆ alkyl),—NH(CH₂)_(n)—(C₃-C₆ cycloalkyl), —NH(CH₂)_(n)-phenyl, —NH(CH₂)_(n)-(3-6membered heterocyclyl), —NH(CH₂)_(n)-(5-6 membered heteroaryl), —N(C₁-C₄alkyl)₂, —N[(CH₂)_(n)—(C₃-C₆cycloalkyl)]₂, —N[(CH₂)_(n)-phenyl]₂,—N[(CH₂)_(n)-(3-6 membered heterocyclyl)]₂, —N[(CH₂)_(n)-(5-6 memberedhetero aryl)]₂, OH, —O(C₁-C₆ alkyl), —O(CH₂)_(n)—(C₃-C₆ cycloalkyl),—O(CH₂)_(n)-phenyl, —O(CH₂)_(n)-(3-6 membered heterocyclyl) or—O(CH₂)_(n)-(5-6 membered heteroaryl).
 10. The compound of claim 1,wherein each R^(a), R^(b) and R^(c) is independently H, C₁-C₄ alkyl,C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, —(C₁-C₂ alkylene)-(C₃-C₆cycloalkyl), 3-6 membered heterocyclyl, —(C₁-C₂ alkylene)-(3-6 memberedheterocyclyl), phenyl, —(C₁-C₂ alkylene)-phenyl, 5-6 membered heteroarylor —(C₁-C₂ alkylene)-(5-6 membered heteroaryl), or R^(a) and R^(b) takentogether with the nitrogen atom to which they are attached form a 3-6membered heterocyclyl group, wherein each of the above substituents isoptionally independently substituted by 1, 2 or 3 substitutentsindependently selected from F, Cl, CN, N₃, OH, NH₂, C₁-C₄ alkyl, C₁-C₄haloalkyl, C₁-C₄ alkoxy and C₁-C₄ alkylamino
 11. The compound of claim1, wherein Z is:

or a stereoisomer thereof, wherein each X, X′, X² and X³ isindependently CH₂, NH or O, with the proviso that when X² is O, X³ isnot O; and wherein Z is optionally substituted by 1, 2 or 3R² groups.12. The compound of claim 1, wherein A is:

wherein A is optionally substituted by 1, 2 or 3R⁴ groups.
 13. Thecompound of claim 1, wherein Z¹ is H, methyl, ethyl, n-propyl, isopropylor cyclopropyl.
 14. The compound of claim 1, wherein R¹ is H, F, Cl, CN,N₃, C₁-C₄ alkyl, C₂-C₄alkenyl, C₂-C₄ alkynyl, C₁-C₄ haloalkyl, C₁-C₄alkoxyl, C₃-C₆ cycloalkyl, 3-6 membered heterocyclyl,—(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b), —C(═O)R⁵,—(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b) or —S(═O)₂NR^(a)R^(b), wherein R¹ isoptionally substituted by 1, 2 or 3R⁸ groups.
 15. The compound of claim1, wherein each R² is independently H, F, Cl, CN, N₃, NO₂, OH, NH₂,—C(═O)CH₂CN, C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₁-C₄ haloalkyl,C₁-C₄ alkoxy, C₃-C₆ cycloalkyl, phenyl, 3-6 membered heterocyclyl, 5-6membered heteroaryl, —(CR⁶R⁷)_(n)—OR^(c), —(CR⁶R⁷)_(n)—NR^(a)R^(b),—C(═O)R⁵, —S(═O)₂R⁵, —O(CR⁶R⁷)_(n)—R⁵, —O(CR⁶R⁷)_(n)—OR^(c),—N(R^(c))C(═O)R⁵, —(CR⁶R⁷)_(n)C(═O)NR^(a)R^(b), —N(R⁹S(═O)_(m)R⁵ or—S(═O)₂NR^(a)R^(b), wherein each R² is optionally independentlysubstituted by 1, 2 or 3R⁸ groups.
 16. The compound of claim 1, whereineach R⁵ is independently H, C₁-C₄ alkyl, C₁-C₄ haloalkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, C₃-C₆ cycloalkyl, phenyl, 3-6 membered heterocyclyl or5-6 membered heteroaryl, wherein each R⁵ is optionally independentlysubstituted by 1, 2 or 3R⁸ groups.
 17. The compound of claim 1 havingone of the following structures:

or a stereoisomer, a tautomer, an N-oxide, a solvate, or apharmaceutically acceptable salt thereof.
 18. A pharmaceuticalcomposition comprising the compound of claim 1, and a pharmaceuticallyacceptable excipient, carrier, adjuvant, vehicle or a combinationthereof.
 19. The pharmaceutical composition of claim 18 furthercomprising a therapeutic agent selected from the group consisting ofchemotherapeutic agents, anti-proliferative agents, phosphodiesterase 4(PDE4) inhibitors, β₂-adrenoreceptor agonists, corticosteroids,non-steroidal GR agonists, anticholinergic agents, antihistamines,anti-inflammatory agents, immunosuppressants, immunomodulators, agentsfor treating atherosclerosis, agents for treating pulmonary fibrosis andcombinations thereof.
 20. A method of preventing, treating or lesseningthe severity of a protein kinase-mediated disease in a patient byadministering to the patient with the compound of claim
 1. 21. Themethod of claim 20, wherein the protein kinase-mediated disease is aJAK-mediated disease, a FLT3-mediated disease, an Aurora-mediateddisease, a proliferative disease, an autoimmune disease, an allergicdisease, an inflammatory disease, a transplantation rejection, cancer,polycythemia vera, essential thrombocytosis, myelofibrosis, chronicmyelogenous leukemia (CML), acute myeloid leukemia (AML), acutelymphocytic leukemia (ALL), chronic obstruction pulmonary disease(COPD), asthma, systemic lupus erythematosis, cutaneous lupuserythematosis, lupus nephritis, dermatomyositis, Sjogren's syndrome,psoriasis, type I diabetes mellitus, allergic airway disease, sinusitis,eczema, hives, food allergies, allergies to insect venom, inflammatorybowel syndrome, Chron's disease, rheumatoid arthritis, juvenilearthritis, psoriatic arthritis, organ transplant rejection, tissuetransplant rejection or cell transplant rejection.
 22. A method ofpreventing, treating or lessening the severity of a proteinkinase-mediated disease in a patient by administering to the patientwith the pharmaceutical composition of claim
 18. 23. The method of claim22, wherein the protein kinase-mediated disease is a JAK-mediateddisease, a FLT3-mediated disease, an Aurora-mediated disease, aproliferative disease, an autoimmune disease, an allergic disease, aninflammatory disease, a transplantation rejection, cancer, polycythemiavera, essential thrombocytosis, myelofibrosis, chronic myelogenousleukemia (CML), acute myeloid leukemia (AML), acute lymphocytic leukemia(ALL), chronic obstruction pulmonary disease (COPD), asthma, systemiclupus erythematosis, cutaneous lupus erythematosis, lupus nephritis,dermatomyositis, Sjogren's syndrome, psoriasis, type I diabetesmellitus, allergic airway disease, sinusitis, eczema, hives, foodallergies, allergies to insect venom, inflammatory bowel syndrome,Chron's disease, rheumatoid arthritis, juvenile arthritis, psoriaticarthritis, organ transplant rejection, tissue transplant rejection orcell transplant rejection.
 24. A method of modulating the activity of aprotein kinase with the compound of claim
 1. 25. The method of claim 24,wherein the protein kinase is JAK kinase, FLT3 kinase, Aurora kinase ora combination thereof.
 26. A method of modulating the activity of aprotein kinase with the pharmaceutical composition of claim
 18. 27. Themethod of claim 26, wherein the protein kinase is JAK kinase, FLT3kinase, Aurora kinase or a combination thereof.